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Phytochemical analysis of Stachys iva: Discovering the optimal extract conditions and its bioactive compounds

  • Aikaterina Vantsioti ORCID logo , Ioannis Makrygiannis ORCID logo , Vassilis Athanasiadis ORCID logo EMAIL logo , Stavros I. Lalas ORCID logo and Paraskevi Mitlianga ORCID logo
Published/Copyright: February 19, 2025
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Open Life Sciences
From the journal Open Life Sciences Volume 20 Issue 1

Abstract

The Lamiaceae family is one of the widest plant families among Greek flora, consisting of a great variety of species, with the genus Stachys being one of its largest representatives, spread to most continents. The genus Stachys is also known for its beneficial properties and has been used for years as a traditional remedy for healing various health conditions. Stachys iva, an endemic plant in the Kozani Regional unit, has also been consumed as an infusion by locals and is reported to relieve common cold symptoms, have antimicrobial properties, and contribute to normalizing blood glucose levels. The present study aimed to identify the chemical compounds (such as phenolic acids, flavonoids, and phenylethanoid glycosides) responsible for the herb’s pharmacological properties and determine the optimal extraction conditions to gather an extract with high therapeutic value without solvent and energy waste. Experiments conducted proved that extracting by simple stirring with deionized water for 75 min at 80°C is the best option. In contrast, the extract’s total polyphenol content was determined, and the compounds were identified by high-performance liquid chromatography analysis. In addition, other methods were utilized (e.g., ferric-reducing antioxidant power assay and 2,2-diphenyl-1-picrylhydrazyl antiradical activity assay) to reveal potent antioxidant, anti-hydrogen peroxide, and anti-inflammatory activity, while the correlation between these properties and extraction conditions was also examined.

Graphical Abstract

Keywords: Stachys ; optimization; polyphenols; antioxidant; anti-inflammatory

1 Introduction

Genera of the Lamiaceae family are widely known for their therapeutic properties and the variety of traditional uses as remedies over the ages. Since ancient times, different Lamiaceae species, conditioned as herbal infusions, edible greens, and extracts, have been mostly consumed as antioxidant, anti-inflammatory, antimicrobial, and analgesic herbal medicines [1,2]. Specific examples of genera from the Lamiaceae family known for their therapeutic properties include Lavandula (lavender) for its calming effects, Mentha (mint) for digestive health, and Salvia (sage) for its anti-inflammatory properties. All the above properties are also common among the species of genus Stachys, one of the largest representatives of the Lamiaceae family as it includes more than 300 species, spread around the Mediterranean area, Asia, America, and southern Africa [3]. The phytochemical analysis conducted on several Stachys species has scientifically verified the empirical use of these herbs as a treatment for different conditions. The polyphenolic compounds which have been isolated and identified are related to the inhibition of lipid peroxidation and result in antioxidant and anti-inflammatory activity [4,5,6]. In addition, flavonoids, iridoids, and glucosides seem to enhance the anti-inflammatory activity. Fatty acids, alkaloids, triterpenes, lignans, and secondary metabolites have also been isolated from different Stachys species [2,7,8]. The numerous pharmacological activities of the latter are also significant as – except for those mentioned above – some species have been efficiently used as antibacterial agents [9,10,11,12], as well as treatment for polycystic ovary syndrome and as potent hepatoprotective, as well as neuroprotective agents against Alzheimer’s disease. Extracts have been additionally examined for their anti-proliferative cytotoxic and antidiabetic properties, proving once again the plethora of pharmacological activities as a potent alternative approach to diabetes and cancer treatment [2,3,5,8,10,13,14].

Despite the plethora of Stachys’ beneficial properties, the potential side effects or even the possibility of interaction among its extracts and other medications should be considered before consuming its infusions. Even though there is not adequate information about Stachys iva toxicity or contraindications – as it has not been thoroughly studied before – some Stachys species are mentioned to be emmenagogue and uterotonic while others cause hypotension [15] or even are toxic damaging liver and renal tissue [16]. In any case, as the above data are derived from experiments in rats, pregnant women and individuals with chronic health issues consuming medicaments should not better consume any herbal remedies without a specialist’s advice due to lack of evidence about potential adverse effects.

Polyphenolic compounds found in abundance in Stachys extracts are more likely to be responsible for their therapeutic properties. These compounds can be found in many fruits, vegetables, and plants, being associated with their beneficial properties and some of their characteristics such as their color, flavor, and odor [17]. Chemically, they contain phenolic rings combined with various structures which are responsible for their classification as well as for their properties. Phenolic acids, flavonoids, lignans, and stilbenes are the main groups of chemical compounds classified as polyphenols [18].

Several studies have been conducted to determine how polyphenol consumption improves our health [19,20]. Primarily, polyphenols are considered to be a natural “weapon” against inflammation. Even inflammation is a natural response of our immune system after tissue damage or infection as an attempt to restore the primer healthy condition, it is characterized by unpleasant symptoms such as redness, pain, swelling, and heat leading, if not suppressed on time, to loss of tissues or even organ’s function. Since the pathophysiology of inflammation is complicated, involving immune cell responses (mainly by cytokines) and a plethora of biochemical molecules that trigger the procedure (such as prostaglandins, cyclooxygenase, and TNFa), its suppression requires a kind of intervention in this “mechanism” [21].

Polyphenolic compounds are also characterized as natural antioxidants, providing oxidative stability to the plant and scavenging free radicals when consumed [17,20]. Even though reactive oxygen species (ROS) are necessary for signal transmission involved in crucial biological functions, their excessive production results in bonding with macromolecules, causing DNA damage, and mutations and affecting normal functions in general [22]. As a result, chronic exposure to oxidative stress due to free radical formation is the main “suspect” for causing cancer, inflammation, as well as degenerative and circulatory system conditions [23,24,25].

Summarizing, polyphenols seem to have a protective role in inflammatory (chronic) conditions, neurodegenerative diseases, and many other consequences of aging in general, whereas several epidemiological studies have co-related the polyphenolic compounds consumption with the reduction of coronary artery disease risk and glucose levels in the blood as well [26]. Even though Stachys species have been thoroughly studied, some endemic species such as Stachys iva are still “unexplored.” S. iva, known as “fluffy mountain tea,” grows in Northern Greece, especially in the mountains of Kozani Regional unit (Western Macedonia), at an altitude of approximately 600 m [27]. It is a perennial herb with a hairy four-angled stem and opposite narrow leaves. Flowers occur in summer, and the corollas are yellow, sympetalous, and two-lipped [28]. A typical S. iva plant with its leaves is illustrated in Figure 1.

Figure 1 A typical Stachys iva plant.
Figure 1

A typical Stachys iva plant.

As S. iva has been traditionally used as a herbal infusion instead of Sideritis spp. due to its antibacterial properties as well as a remedy for the relief of common cold symptoms, the present study aims to identify the chemical compounds that could be responsible for these therapeutic properties. Regarding the identification of the active compounds, high-performance liquid chromatography (HPLC) has been utilized, combined with other conventional experimental procedures. Specifically, Folin–Ciocalteu and ferric-reducing antioxidant power (FRAP) assay, in addition to 2,2-diphenyl-1-picrylhydrazyl (DPPH) antiradical activity assay and hydrogen peroxide scavenging assay, were conducted for the determination of the total phenolic compounds and antioxidant/antiradical capacity of the extract. The anti-inflammatory activity of the S. iva extract has also been assessed in vitro as an attempt to experimentally justify some of the plant’s pharmacological properties.

An additional purpose of this study is to detect the optimal extraction conditions of S. iva in order to investigate which temperature–duration combination leads to the most enriched, in bioactive compounds, extract. The optimization also has an environmentally friendly aspect, as it aims to the reduction of energy and solvent consumption. Moreover, using water as solvent through the process does not involve harmful organic solvents which are harmful to the environment and may provide extracts unsuitable for consumption by humans. To the best of our knowledge, there are no other comprehensive phytochemical analyses of S. iva available in the literature. Therefore, our study represents a novel contribution to the understanding of this species’ bioactive compounds.

2 Materials and methods

2.1 Chemicals and reagents

All the experiments were conducted by using mainly deionized water as solvent except for solvents used in chromatography (water, formic acid, and acetonitrile) that were HPLC grade. Neochlorogenic acid, chlorogenic acid, rutin, verbascoside, narirutin, myricetin, and rosmarinic acid were purchased from Sigma-Aldrich (Darmstadt, Germany) as chemical standards and used in HPLC for determination of polyphenolic compounds. Deionized water was chosen as the extraction solvent due to its safety, cost-effectiveness, and ability to extract a wide range of polyphenolic compounds.

Hydrochloric acid, phosphate buffer solution, methanol, l-ascorbic acid, trichloroacetic acid, aluminum chloride, 2,4,6-tris(2-pyridyl)-s-triazine (TPTZ), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) were also purchased from Sigma-Aldrich (Darmstadt, Germany). The Folin–Ciocalteu reagent, ethanol, and gallic acid were obtained from Panreac Co. (Barcelona, Spain). Hydrogen peroxide (35% v/v) was purchased from Chemco (Malsch, Germany), iron(iii) chloride from Merck (Darmstadt, Germany), and anhydrous sodium carbonate from Penta (Prague, Czech Republic). Egg albumin used for the detection of anti-inflammatory action was provided by eggs purchased from the local market.

2.2 Plant material

Plant materials (leaves, stems, and flowers) were collected in the blooming stage in June 2023 from Polyrrachos village (at 40°08′23.0″N and 21°57'12.0″E, based on Google Earth version 9.185.0.0) in Kozani, Municipality of Western Macedonia Greece, at about 630 m altitude. The microclimate of the area is characterized by warm summers, cool autumns and springs, and cold winters, wetter than other regions of the municipality affected by the Polyphytos Lake nearby.

After collection, the aerial parts of S. iva were weighed and dried for 30 days at room temperature in a shaded room without excessed humidity until their weight stabilized, indicating complete loss of water, and were subjected to extraction processes. The aerial parts were ground after drying, and the particle size of the dried plant material was determined.

2.3 Plant extraction with response surface methodology (RSM)

In order to maximize the efficiency of extracting polyphenols and assessing antioxidant and anti-inflammatory activity from medicinal plant extracts, the RSM technique was utilized. First, the optimal temperature and extraction time were studied. To do this, 0.4 g of the S. iva plant was accurately weighed (Kern PLS 3100-2F, Kern & Sohn GmbH, Balingen, Germany) using 20 mL of deionized water. As shown in Table 1, the experiments were conducted in a temperature range of 20–80°C and the time of extraction ranged from 30 to 90 min. The experiment that used a Main Effects Screening design with a major impact screening arrangement served as the basis for the optimization procedure. There were 10 design points in the experiment. The experimental design resulted in the creation of five levels of process variables. Analysis of variance (ANOVA) and summary-of-fit tests were used to evaluate the overall model significance, as indicated by the R 2 and p-values, and the significance of the model coefficient, as indicated by the equation, with a minimum level of 95% confidence. After the 500-rpm stirring extraction process was finished, the samples were centrifuged at 10,000 × g for 10 min at room temperature using a NEYA 16R Remi Elektrotechnik Ltd. (Palghar, India). The supernatants were then gathered and kept in storage at −40°C.

Table 1

Actual and coded levels of the independent variables were used to optimize the extraction process using the Main Effects Screening design

Independent variables Code units Coded variable level
1 2 3 4 5
Temperature (T, °C) X 1 20 35 50 65 80
Time (t, min) X 2 30 45 60 75 90

Additionally, a quadratic (second-order) polynomial model was used to predict the response variable as a function of the investigated independent components, as shown by equation (1):

(1) Y k = β 0 + ∑ i = 1 2 β i X i + ∑ i = 1 2 β ii X i 2 + ∑ i = 1 2 ∑ j = i + 1 3 β ij X i X j .

The predicted response variable is denoted as Y k , while the independent variables are X i and X j . The intercept and regression coefficients for the linear, quadratic, and interaction terms of the model are denoted as β 0, β i , β ii , and β ij , respectively.

The RSM was used to identify the largest peak region and evaluate the impact of a significant independent variable on the response [29]. To visually depict the model equation, the creation of a three-dimensional surface response graph took place.

2.4 Other extraction techniques

In order to confirm that water is the optimal extraction solvent, other solvents were used for extraction purposes, such as ethanol as an alternative solvent. Specifically, 2 g of the dried S. iva sample were extracted in 100 mL ethanol as well as in ethanol with water mixture (60:40 v/v) at 50°C for 30 min. In contrast, two more plant samples of the same weight were extracted in deionized water at 100°C for 5 min and at 50°C for 30 min in order to collect comparable results.

2.5 Polyphenol determination

2.5.1 Total polyphenol content (TPC)

A previously established methodology [30] was applied to determine TPC. Briefly, 100 μL of the properly diluted sample extract was mixed with 100 μL of Folin–Ciocalteu reagent, and after 2 min, 800 μL of 5% w/v aqueous sodium carbonate solution was added. The mixture was incubated at 40°C for 20 min and the absorbance was recorded at 740 nm in a Shimadzu UV-1700 PharmaSpec Spectrophotometer (Kyoto, Japan). The TPC (C TP) was calculated from a gallic acid calibration curve (10–80 mg/L). TPC was determined as mg gallic acid equivalents (GAE) per g of dry weight (dw), using the following equation:

(2) TPC ( mg GAE / g dw ) = C TP × V w ,

where the volume of the extraction medium is indicated with V (expressed in L) and the dry weight of the sample as w (expressed in g).

2.5.2 HPLC quantification of polyphenolic compounds

HPLC was used to detect and quantify individual polyphenols from the sample extracts, as established in our previous research [30]. A Shimadzu CBM-20A liquid chromatograph and a Shimadzu SPD-M20A diode array detector (DAD) (both purchased by Shimadzu Europa GmbH, Duisburg, Germany) were employed for the analysis of medicinal plant extracts. The compounds were separated into a Phenomenex Luna C18(2) column from Phenomenex Inc. (Torrance, CA, USA), kept at 40°C (100 Å, 5 μm, 4.6 mm × 250 mm). The mobile phase included 0.5% aqueous formic acid (A) and 0.5% formic acid in acetonitrile/water (3:2) (B). The gradient program required: initially from 0 to 40% B, then to 50% B in 10 min, to 70% B in another 10 min, and then constant for 10 min. The flow rate of the mobile phase was set at 1 mL/min. The compounds were identified at 320 nm by comparing the absorbance spectrum and retention time to those of pure standards and then quantified through calibration curves (0–50 μg/mL).

2.6 Antioxidant capacity of the extracts

2.6.1 FRAP assay

An established technique by Shehata et al. [31] was used for the evaluation of FRAP. In a 1.5-mL Eppendorf tube, 50 μL of the properly diluted sample extract was mixed with 50 μL of FeCl3 solution (4 mM in 0.05 M HCl). The mixture was incubated for 30 min at 37°C, with 900 μL of TPTZ solution (1 mM in 0.05 M HCl) being immediately added right after, and the absorbance was measured after 5 min at 620 nm. The ferric-reducing power (P R) was calculated using an ascorbic acid calibration curve (C AA) in 0.05 M HCl with ranging values (50–500 μM). The P R was calculated as μmol of ascorbic acid equivalents (AAE) per g of dw, using equation (3):

(3) P R ( μ mol AAE / g d w ) = C AA × V w ,

where V is represented (in L) as the entire volume of the extraction medium and w (in g) represents the dried weight of the material.

2.6.2 DPPH• antiradical activity assay

The extracted polyphenols from the dried material were evaluated for their antiradical activity (A AR) using a slightly modified DPPH˙ method, as previously established by Shehata et al. [31]. In brief, 25 μL of the properly diluted sample extract was mixed with a quantity of 975 μL of a 100 μM DPPH˙ solution in methanol, with the solution being kept at room temperature for 30 min in the dark right after. The absorbance was measured at 515 nm. Moreover, a blank sample was used instead of the sample, including DPPH˙ solution and methanol, with the absorbance immediately being measured. To calculate the percentage of scavenging, equation (4) was employed:

(4) % Scavenging = A control − A sample A control × 100 .

An ascorbic acid calibration curve (C AA, 100–1,000 μmol/L in methanol) in equation (5) was used to evaluate antiradical activity (A AR), which was expressed as μmol AAE per g of dw:

(5) A AR ( μ mol AAE / g d w ) = C AA × V w ,

where V is represented (in L) as the entire volume of the extraction medium and w (in g) represents the dried weight of the material.

2.6.3 Hydrogen peroxide (H2O2) scavenging assay

A previous method [32] was applied for the H2O2 scavenging assay. A quantity of 400 μL of the properly diluted sample extract and 600 μL of an H2O2 solution (40 mM, made in phosphate buffer, pH 7.4) was added into an Eppendorf tube. The absorbance was recorded right after 10 min at 230 nm. The scavenging capacity of the H2O2 was expressed as follows:

(6) % Scavenging of H 2 O 2 = A 0 − ( A − A c ) A 0 × 100 ,

where the absorbances of the blank solution, the extract solution in the absence of hydrogen peroxide, and the sample are denoted by A 0, A c, and A, respectively.

The concentration of ascorbic acid ranged in the calibration curve (C AA, 50–500 μmol/L in 0.05 M HCl) and the following equation (7) was used to determine the anti-hydrogen peroxide activity (A AHP) as μmol AAE per g of dw:

(7) A AHP ( μ mol AAE / g d w ) = C AA × V w ,

where V denotes the volume of the extraction medium (in L), and w is the dry weight of the sample.

2.7 Assessment of in vitro anti-inflammatory activity

The in vitro evaluation of the anti-inflammatory properties of the S. iva extracts was conducted using the albumin denaturation assay [33]. Briefly, a mixture containing egg albumin and PBS (pH = 6.4) with a 0.1:1.4 mL ratio, or 6.67% v/v (mixture A), respectively, was prepared. Then, 400 μL of the properly diluted sample extract or standard were mixed with 600 μL of mixture A in a 1.5 mL Eppendorf tube, and then, the tubes were incubated at 37°C for 15 min. Afterwards, the mixture was heated at 70°C for 5 min. The absorbance was then recorded at 660 nm. This wavelength was chosen to minimize interference from the extract’s color or turbidity. To determine the % inhibition of protein denaturation, the following equation (8) was used:

(8) % Inhibition = Absorbance sample Absorbance control − 1 × 100 .

By measuring the absorbance of the control (without extract) against that of the sample (with extract), we can assess the degree to which the extract prevents protein denaturation.

2.8 Statistical analysis

The statistical analysis was applied to the RSM through Main Effects Screening design and distribution analysis, using the JMP® Pro 16 program (SAS, Cary, NC, USA). The extraction processes were carried out at least twice for every batch of plant extract, and the quantitative analysis was carried out in triplicate. The Kolmogorov–Smirnov test was employed to verify data normality. To detect statistically significant differences, a one-way ANOVA was conducted, succeeded by a post hoc Tukey HSD (honestly significant difference) test using the Tukey–Kramer method. The means and standard deviations of the results are used to demonstrate them. Using JMP® Pro 16 software, Pareto plot analysis, principal component analysis (PCA), and multivariate correlation analysis (MCA) were carried out.

3 Results and discussion

3.1 Optimization of extraction conditions

Even though extraction is considered to be a common laboratory technique, it may result in being energy consuming, requiring also a significant amount of solvents, as each plant has a divergent abundance of bioactive compounds with different chemical properties. Supposing that cost-effectiveness is not taken into consideration, the majority of the organic solvents being used are harmful to the environment. Moreover, it may be necessary for the plant material to be extracted with many different solvents in order to conclude which of them and under which conditions is able to extract as many substances as possible, providing the researcher with an extraction that may lead to significant conclusions about the plant’s potential properties [34]. Additionally, the extraction duration and temperature are crucial parameters that define the energy consumption during the procedure. Obviously, the detection of optimal extraction conditions is necessary as it enhances the efficiency of the procedure, and utilizing water as solvent seems to be the most suitable choice, as it is approachable, affordable, and without environmental impact [35]. Investigating the effect of temperature and duration on the extraction process has improved our understanding of optimizing the yield and efficacy of polyphenolic compounds. This knowledge is crucial for devising more effective extraction techniques that ensure the preservation and maximization of bioactive compounds.

Taking all the above into consideration, as well as the predicted antioxidant activity and TPC of the extract, we came up with a correlation between the extraction conditions and the extract’s properties using the RSM approach, as presented in Tables 2 and 3, as an attempt to investigate the optimal design point. Obviously, extracting the plant material for 75 min at 80°C (design point-DP 5) seems to provide us with an extract rich in polyphenolic compounds and with enhanced antioxidant potency, as it is demonstrated in Figure 2, where the predicted response referring to parameters of DP 5 presenting as statistically significant. The optimal temperature was quite anticipated, as there are references documenting that the highest polyphenolic substance recovery is conducted under 50–80°C [36]. Also, the three-dimensional plot for TPC can be seen in Figure 3.

Table 2

Experimental findings for the two independent variables under investigation and the dependent variable’s response to TPC

Design point Independent variables Response TPC (mg GAE/g dw)
X 1 (T, °C) X 2 (t, min) Actual Predicted
1 1 (20) 2 (45) 9.02 ± 0.62 10.27
2 2 (35) 2 (45) 7.69 ± 0.43 7.76
3 3 (50) 3 (60) 11.33 ± 0.56 10.39
4 4 (65) 4 (75) 15.75 ± 1.04 15.46
5 5 (80) 4 (75) 31.13 ± 2.18 29.52
6 1 (20) 1 (30) 8.96 ± 0.34 7.58
7 2 (35) 1 (30) 10.92 ± 0.29 11.57
8 3 (50) 5 (90) 18.12 ± 1.20 17.70
9 4 (65) 3 (60) 19.72 ± 0.67 21.10
10 5 (80) 5 (90) 21.67 ± 1.13 22.97
Table 3

Coded values of the two independent variables under investigation and the actual values of antioxidant and anti-inflammatory activity assays

Design point Independent variables Responses
X 1 (T, °C) X 2 (t, min) FRAP (μmol AAE/g dw) DPPH (μmol AAE/g dw) Anti-hydrogen peroxide activity (μmol AAE/g dw) Anti-inflammatory activity (%)
1 1 (20) 2 (45) 47.02 ± 3.53 17.81 ± 1.26 129.78 ± 5.84 42.53 ± 2.64
2 2 (35) 2 (45) 41.55 ± 2.99 14.03 ± 0.86 76.86 ± 1.77 42.30 ± 3.13
3 3 (50) 3 (60) 80.05 ± 1.68 32.92 ± 1.35 192.42 ± 14.24 44.50 ± 2.18
4 4 (65) 4 (75) 94.69 ± 5.40 59.18 ± 3.08 301.86 ± 14.79 56.02 ± 3.53
5 5 (80) 4 (75) 182.85 ± 11.15 95.83 ± 5.17 459.36 ± 31.70 72.53 ± 4.71
6 1 (20) 1 (30) 51.04 ± 3.67 16.29 ± 0.46 82.44 ± 3.38 47.61 ± 3.52
7 2 (35) 1 (30) 60.73 ± 3.16 22.41 ± 1.43 127.80 ± 6.77 55.69 ± 1.28
8 3 (50) 5 (90) 98.50 ± 3.25 47.72 ± 2.72 270.18 ± 7.57 55.09 ± 3.97
9 4 (65) 3 (60) 107.31 ± 3.97 49.69 ± 1.94 280.98 ± 16.02 60.83 ± 3.35
10 5 (80) 5 (90) 131.41 ± 6.44 58.88 ± 3.36 348.84 ± 10.12 64.17 ± 4.17
Figure 2 Plot (a) displays the actual versus the predicted response (TPC, mg GAE/g dw) for the optimization of extraction of S. iva plant performed with water solutions. The inset tables provide statistics related to the evaluation of the resulting model. Values with color and asterisk are statistically significant. The desirability function for the optimization of extraction of S. iva performed with water solutions is displayed in Plot (b).
Figure 2

Plot (a) displays the actual versus the predicted response (TPC, mg GAE/g dw) for the optimization of extraction of S. iva plant performed with water solutions. The inset tables provide statistics related to the evaluation of the resulting model. Values with color and asterisk are statistically significant. The desirability function for the optimization of extraction of S. iva performed with water solutions is displayed in Plot (b).

Figure 3 Three-dimensional graph depicting the covariation of X 1 (T, °C) and X 2 (t, min) and the effect of the process variables considered on the response (TPC, mg GAE/g dw) for the optimization of extraction of S. iva performed in water solutions.
Figure 3

Three-dimensional graph depicting the covariation of X 1 (T, °C) and X 2 (t, min) and the effect of the process variables considered on the response (TPC, mg GAE/g dw) for the optimization of extraction of S. iva performed in water solutions.

The desirability function was used in the optimization process to combine multiple response variables into a single composite response. Each response variable was transformed into a desirability value ranging from 0 (completely undesirable) to 1 (fully desirable). The overall desirability was calculated as the geometric mean of the individual desirability values, providing a single metric that reflects the optimal conditions for all response variables. This approach allows for the simultaneous optimization of multiple criteria, ensuring a balanced and comprehensive evaluation of the extraction conditions.

The TPC was expressed in milligrams of gallic acid equivalents per gram of dry weight (mg GAE/g dw). This unit standardizes the phenolic content based on a gallic acid calibration curve, allowing for comparison across different studies and samples. The antioxidant capacity measured by assays such as FRAP and DPPH was expressed in micromoles of ascorbic acid equivalents per gram of dry weight (µmol AAE/g dw). This unit standardizes the antioxidant activity based on an ascorbic acid calibration curve.

The ferric-reducing antioxidant power (FRAP) assay, commonly utilized to assess the antioxidant capacity of different samples, presents multiple limitations. Its high sensitivity to pH fluctuations, especially it is performed at an acidic pH of approximately 3.6, may not provide an accurate measure of antioxidant capacity at physiological pH. Moreover, temperature changes can affect the outcomes, with elevated temperatures possibly causing an overestimation and reduced temperatures an underestimation. The FRAP assay is designed to measure the reducing power of antioxidants, which may not reflect all antioxidant activities, such as radical scavenging or metal chelation. This limitation restricts its capacity for a complete assessment. Additionally, the assay can be affected by other reducing agents present in the sample, potentially leading to erroneous outcomes. Nevertheless, the FRAP assay is valued for its simplicity, rapidity, cost-efficiency, and consistent reproducibility when standard conditions are applied. Ascorbic acid is often employed as a benchmark for its notable antioxidant properties; however, it might not adequately reflect the intricate antioxidant activities of plant extracts. Employing alternative methods, such as a blend of standards like Trolox, or integrating the FRAP assay with other antioxidant evaluations (for instance, DPPH, ABTS, ORAC) could offer a more comprehensive evaluation of antioxidant potential by encompassing various mechanisms of action.

The second-order polynomial equations obtained for the responses were expressed as follows:

(9) TPC = 9.83 − 4.29 X 1 + 0.82 X 2 + 4.93 X 1 2 + 2.79 X 2 2 − 6.5 X 1 X 2 ,

(10) FRAP = 50.19 − 19.58 X 1 + 5.14 X 2 + 22.88 X 1 2 + 11.88 X 2 2 − 28.27 X 1 X 2 ,

(11) DPPH = 6.75 − 8.96 X 1 + 10.54 X 2 + 11.95 X 1 2 + 5.28 X 2 2 − 15.06 X 1 X 2 ,

(12) Anti-hydrogen peroxide activity = 49.87 − 47.11 X 1 + 56.22 X 2 + 54.34 X 1 2 + 23.84 X 2 2 − 66.99 X 1 X 2 ,

(13) Anti-inflammatory activity = 55.25 − 0.23 X 1 − 10.95 X 2 + 5.75 X 1 2 + 4.99 X 2 2 − 8.09 X 1 X 2 ,

Depending on ANOVA analysis results, we may also evaluate the responses of the extract, as an attempt to assess its potent properties. Apparently, all the responses of the conducted experiments seem to be statistically significant, indicating the pharmacological properties of S. iva plant extract. Moreover, according to the RMSE, total phenolic content and anti-inflammatory activity appear to be closer to the actual values, so we anticipated these properties to be experimentally proved, as demonstrated in Table 4.

Table 4

ANOVA analysis of the responses

Responses F-ratio p-value R 2 Adjusted R 2 RMSE
TPC 34.24 0.0022 0.9772 0.9486 1.67
FRAP 15.65 0.0099 0.9514 0.8906 14.54
DPPH 8.57 0.0292 0.9146 0.8078 11.38
Anti-hydrogen peroxide activity 22.03 0.0052 0.9650 0.9212 35.41
Anti-inflammatory activity 46.11 0.0013 0.9829 0.9616 1.96

RMSE means root mean square error.

3.2 Impact of extraction parameters on assays through Pareto plot analysis

In Pareto plot analysis, factors are chosen for their contribution to total variance, with the most influential ones accounting for the largest percentage of variance. In order to examine the positive or negative impact of extraction parameters on the extract properties, we conducted a Pareto plot analysis including results from all the experimental assays that took place. Observing the plots (Figure 4), we may conclude to the fact that the combination of temperature and time of extraction does not affect either the TPC of the extract or its antioxidant and anti-inflammatory properties, as it is demonstrated to adopt negative or very low values. On the other hand, the temperature itself (factor X 1) mainly contributes to eventually collecting the optimal extract. The temperature not only affects the extract’s quality but also overcomes the critical value’s threshold (gold line) indicating statistical significance. Pareto plot analysis may occasionally simplify the complex interplay between factors too much, which could lead to a misinterpretation of the results.

Figure 4 Pareto plots of transformed estimates for TPC (a), FRAP (b), DPPH (c), anti-hydrogen peroxide activity (d), and anti-inflammatory activity (e) assays. A reference gold line is drawn on the plot to indicate the level of significance (p < 0.05).
Figure 4

Pareto plots of transformed estimates for TPC (a), FRAP (b), DPPH (c), anti-hydrogen peroxide activity (d), and anti-inflammatory activity (e) assays. A reference gold line is drawn on the plot to indicate the level of significance (p < 0.05).

3.3 Optimal extraction conditions

Attempting to optimize the extraction conditions furthermore, partial least squares (PLS) analysis was used for the assessment of the extraction parameters impact, as depicted in Figure 5. Taking into consideration that desirability equals 0.8496, variables estimated over this point seem to have a significant contribution. As demonstrated in the plot, temperature (X 1 variable) affects the quality of extract, the quantity of polyphenols (TPC), and its antioxidant and anti-inflammatory properties. Specifically, the DP5 (80°C) seems to provide the optimum results. Examining the impact of extraction duration (X 2 variable), it can be noticed that this parameter solely does not determine the extract’s content at the same level comparatively with the temperature. However, extracting S. iva at 80°C for 75 min (DP5) is predicted to be the ideal parameter combination.

Figure 5 Maximum predicted responses and optimum extraction conditions for the dependent variables.
Figure 5

Maximum predicted responses and optimum extraction conditions for the dependent variables.

3.4 PCA and MCA

To obtain a more objective aspect of how the temperature and the duration of the extraction are correlated to the TPC, the antioxidant (FRAP, DPPH, H2O2), and anti-inflammatory assays, a PCA was conducted. In PCA, the selection criteria focus on identifying the principal components that account for the majority of the variance in the data, those with eigenvalues >1. As the parameter’s sign determines whether or not two or more variables are associated, we may conclude once again, according to Figure 6, that the extraction’s temperature positively affects the extract’s properties and its concentration of polyphenolic compounds. Additionally, all the variables were well discriminated, and it was revealed that TPC and FRAP assay, are also positively correlated. On the contrary, the negative sign of extraction’s duration, reveals, as anticipated, that this parameter does not noticeably determine the extract’s composition or its experimental assays measurements. PCA is beneficial for dimensionality reduction but may not consistently capture the non-linear relationships between variables.

Figure 6 PCA for the measured variables. Each X variable is presented with a blue color.
Figure 6

PCA for the measured variables. Each X variable is presented with a blue color.

Multiple correspondence analysis was utilized to discern correlations among various variables, with the most significant factors identified by the highest correlation coefficients. Moreover, taking into consideration the data displayed in Table 5, the high association among all the parameters is also confirmed as they all have positive signs and correlation >0.89. A low correlation, compared to other measured variable pairs, was shown between anti-inflammatory activity and anti-hydrogen peroxide activity (0.8866), in contrast with the highest correlation that appears among the latter and DPPH (0.9858). MCA may be constrained by multicollinearity and the possibility of spurious correlations.

Table 5

Multivariate correlation analysis of measured variables

Responses TPC FRAP DPPH Anti-hydrogen peroxide activity Anti-inflammatory activity
TPC — 0.9691 0.9411 0.9583 0.9419
FRAP — 0.9755 0.9740 0.9177
DPPH — 0.9858 0.8924
Anti-hydrogen peroxide activity — 0.8866
Anti-inflammatory activity —

3.5 Optimal extract and comparison with other extraction techniques

The correlation between the values generated by the second-order polynomial model and those acquired through experimental analysis is determined to be 0.9973, and they show no deviations with the p-value being 0.0002. Comparing data from Table 6, the optimal extract’s responses are similar to the maximum predicted ones and mainly seem more increased than the latter, concluding that extracting at 80°C for 75 min is the ideal condition for S. iva. Extracting in boiling water for 5 min and in water:ethanol mixture (40:60 v/v) at 50°C for 30 min, leads to quite similar and pretty significant experimental responses, except the TPC, but lower than the expecting ones. This mixture was chosen based on its ability to extract a broad range of polyphenolic compounds. The less interesting results that depict significantly decreased compared to the optimal extract are derived if we extract the plant material with pure ethanol or deionized water at 50°C for 30 min. Ethanol was chosen as an alternative solvent due to its effectiveness in extracting polyphenolic compounds and its safety for use in food and pharmaceutical applications.

Table 6

Maximum predicted responses, optimal extract under optimal extraction conditions, and comparison with other extraction techniques

Extracts Responses
TPC (mg GAE/g) FRAP (μmol AAE/g) DPPH (μmol AAE/g) Anti-hydrogen peroxide activity (μmol AAE/g) Anti-inflammatory activity (%)
Maximum predicted response 29.52 ± 3.83a 169.44 ± 33.30a 86.30 ± 26.05a 439.27 ± 81.08a 72.12 ± 4.49a
Optimal extract 31.55 ± 0.79a 185.78 ± 5.57a 95.40 ± 3.91a 410.42 ± 16.42a 70.06 ± 1.47a,b
Boiling water 12.50 ± 0.89c 92.92 ± 2.23b 73.56 ± 4.56a,b 229.14 ± 16.50b 61.74 ± 4.01b
Ethanol 60% v/v 19.99 ± 0.54b 92.88 ± 3.81b 51.74 ± 3.88b 231.84 ± 4.64b na
Ethanol 2.52 ± 0.07d 11.33 ± 0.69c 3.46 ± 0.13c 26.64 ± 0.80c na
Water 10.99 ± 0.31c 32.93 ± 1.75c 19.80 ± 1.05c 63.36 ± 1.96c 49.57 ± 3.32c

Within each column, statistically significant differences (p < 0.05) are denoted with lowercase letters (e.g., a–d); na means not analyzed.

3.6 Analysis of the optimal extract

The experimental assays we conducted, indicated that S. iva extract is abundant in polyphenolic compounds, as we anticipated due to statistical predictions and bibliographic references to other Stachys species. Especially the optimal extract has the ultimate TPC, 31.55 ± 0.79 mg GAE/g dw (actual) and 29.52 ± 3.83 mg GAE/g dw (predicted response), compared to others. The polyphenol compounds as we mentioned are mainly responsible for the plant’s beneficial properties, affecting its antioxidant and anti-inflammatory activity. This is also proved by the fact that all these variables correlate and their values are associated, as results from our data analysis.

The main polyphenolic compounds of the optimal extract were identified and also quantified, data demonstrated in Table 7, by using HPLC-DAD. A representative chromatogram is illustrated in Figure 7, where according to the retention time of each compound and their UV spectrum, phenolic acids such as rosmarinic, neochlorogenic and chlorogenic acid, and flavonoids (rutin, verbascoside, narirutin, myricetin) were identified.

Table 7

Polyphenolic compounds analysis of optimal extract under optimal extraction conditions

Polyphenolic compound Optimal extract (mg/g dw)
Neochlorogenic acid 0.55 ± 0.03
Chlorogenic acid 5.02 ± 0.21
Rutin 0.45 ± 0.02
Verbascoside 0.87 ± 0.06
Narirutin 0.03 ± 0
Myricetin 0.46 ± 0.01
Rosmarinic acid 1.4 ± 0.04
Total identified 8.79 ± 0.38
Figure 7 Representative HPLC chromatogram at 320 nm of optimal extract, demonstrating polyphenolic compounds that were identified. 1: Neochlorogenic acid; 2: chlorogenic acid; 3: rutin; 4: verbascoside; 5: narirutin; 6: myricetin; 7: rosmarinic acid.
Figure 7

Representative HPLC chromatogram at 320 nm of optimal extract, demonstrating polyphenolic compounds that were identified. 1: Neochlorogenic acid; 2: chlorogenic acid; 3: rutin; 4: verbascoside; 5: narirutin; 6: myricetin; 7: rosmarinic acid.

Examining further each compound’s properties we may better understand the traditional use of “fluffy mountain tea’s” extract as a remedy for curing a wide range of health conditions. Initially, referring to rosmarinic acid (RA), many studies have been conducted both in vivo and in vitro, trying to suggest its mechanisms of action against inflammation and oxidative stress. RA’s chemical structure allows the compound to be lipophilic enough to penetrate the cell membrane and protect the cell by scavenging free radicals. Its antioxidant properties protect different organs such as liver and heart enhancing also its potent use as a neuroprotective and anticancer agent. Furthermore, studies are reporting RA’s action against different types of bacteria and fungi as well as against inflammatory conditions, possibly by inhibiting the synthesis of cytokines and suppresses the activation of cyclooxygenase and even of T-leukocytes in autoimmune diseases [37,38,39,40].

Another well-known phenolic acid that is pretty ubiquitous in plant extracts, especially in coffee and tea infusions, and with the highest concentration in our extract is chlorogenic acid (CGA). Despite its action against free radicals and inflammation due to its chemical properties, CGA seems to affect glucose absorption decreasing its concentration in blood circulation. Thus, fat burning increases as an alternative source of energy, leading to both antidiabetic and anti-obesity action. Additionally, clinical trials proved that CGA consumption is associated with attenuation of blood pressure, probably because of the induction of vessel relaxation, antimicrobial activity has also been reported [41,42,43]. As for neochlorogenic acid, that is CGA’s isomer and has similar properties. However, we may highlight that its anti-inflammatory activity derives from the inhibition of lipopolysaccharide (LPS). The latter acts like a stimulant that activates microglia (type of macrophages), causing neuroinflammation and triggering the onset of neurodegenerative diseases [44,45].

Even though phenolic acids concentration in the extract was 6.97 ± 0.28 mg/g dw out of the 8.79 ± 0.38 mg/g dw of totally identified polyphenolic compounds, many flavonoids contribute to S. iva properties and verbascoside (VERB) is the dominant one (0.85 ± 0.06 mg/g dw). VERB is a phenylethanoid glycoside, initially isolated from Verbascum sp. (mullein) but distributed in plenty of other plant species that have also been used as traditional remedies. Studies report that VERB combines anti-inflammatory, antibacterial, and even anti-androgen properties being a potent alternative approach for acne vulgaris treatment. Moreover, preventing lipoprotein oxidation protects cells from oxidative stress while repairs DNA damage caused by the latter. It may also be used as an anti-depressant, acting by triggering monoamine neurotransmitters’ production and increasing their levels and availability, while it seems to appear a protective role against atherosclerosis [46,47,48].

Rutin and myricetin are detected in approximately equal concentrations. They both possess diverse beneficial properties acting in vitro, as the majority of polyphenols, against lipid peroxidation, inflammation, and as radical scavengers [49]. Furthermore, rutin appears to have vasoprotective, anti-hypercholesterolemic, anti-hypertensive, and antiplatelet aggregation effects in vitro and after being administered to rats, while also promoting cancer cell apoptosis as indicated in experiments conducted using cell lines from various cancer types [50,51,52,53]. Highlighting some distinct properties of myricetin, we should mention that there are pieces of evidence about its intervention in diabetes and obesity management in rats [54,55].

Last but not least, even in minimum concentration, the flavonoid narirutin, which is mainly found in citrus fruit, has a plethora of beneficial properties similar to the ones documented for the rest of the extract’s flavonoids [56,57,58,59].

4 Conclusions

The present study aimed to identify the ideal extraction solvent (deionized water and ethanol) and optimal extraction parameters (temperature and duration) to obtain the most enriched extract of Stachys iva, with high therapeutical value. S. iva has been traditionally used as a remedy with diverse pharmacological properties, which this study attempted to validate experimentally. RSM was utilized to identify the optimal extraction conditions, combined with Pareto plot analysis. Additionally, PCA and MCA detected positive correlations between experimental assay responses and extraction conditions, as well as the association among these responses.

Each extract underwent experimental procedures to determine its TPC, antioxidant capacity (FRAP, DPPH, anti-hydrogen peroxide activity), and anti-inflammatory activity. The collected data were compared with predicted values. HPLC analysis identified and quantified polyphenolic compounds, with a total concentration of 8.79 ± 0.38 mg/g dw. Chlorogenic acid was the dominant compound (5.02 ± 0.21 mg/g dw), followed by verbascoside (0.87 ± 0.06 mg/g dw), neochlorogenic acid, rutin, narirutin, myricetin, and rosmarinic acid.

Considering evidence from in vitro/in vivo studies and clinical trials referring to these compounds’ beneficial properties, the traditional use of S. iva extracts as antioxidant, anti-inflammatory, antimicrobial, and antidiabetic agents can be experimentally validated. These compounds also show potential pharmacological properties against neurodegenerative and other diseases, leading to further research on S. iva with promising results.

Future research will explore the use of alternative solvents, such as ethanol, methanol, and acetone, which may enhance the extraction efficiency of bioactive compounds from S. iva. The selection of solvents will be based on their polarity, safety, and environmental impact. Additional response variables such as flavonoid content, tannin content, and specific antioxidant activities (e.g., superoxide dismutase activity) will be investigated to provide a more comprehensive understanding of the bioactive profile of S. iva extracts. Advanced extraction techniques such as supercritical fluid extraction, microwave-assisted extraction, and ultrasound-assisted extraction will be explored to optimize the yield and quality of bioactive compounds. These techniques offer potential benefits in terms of efficiency, selectivity, and environmental sustainability.

Moreover, the investigation of S. iva’s composition of bioactive compounds and the experimental justification of its traditional uses may lead to a well-established use of this herb in products of food, cosmetic, and pharmaceutical industries. S. iva infusions may replace water or other extracts in lotions and creams, providing them with an aqueous phase rich in polyphenols and antioxidants enhancing the skin barrier against free radicals. Due to its abundance in polyphenolic substances, S. iva extract may be added to beverages as flavoring and perhaps as a natural preservative due to its antimicrobial potency, resulting in the production of a functional drink. As for the pharmaceutical industry, even though S. iva has many potential pharmacological properties (antidiabetic, antiproliferative) as mentioned, more research is required in order to ensure the lack of toxicity, severe adverse effects, or even interaction with medications. However, its extract could be included in supplements against the common cold only if the sufficiency of safety data is proven.

Acknowledgments

The authors would like to thank Mr. Christos Papagiannis for collecting Stachys iva and providing us with plant material, as well as for capturing the photograph of the plant in its natural habitat. Also, the authors are grateful for the reviewers’ valuable comments that improved the manuscript.

  1. Funding information: Authors state no funding involved.

  2. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and consented to its submission to the journal, reviewed all the results, and approved the final version of the manuscript. Conceptualization was carried out by A.V. and P.M.; methodology was developed by V.A. and S.I.L.; experimentation was conducted by A.V., I.M., and V.A.; data analysis and figures preparation were performed by A.V., I.M., and V.A.; writing – original draft preparation was done by A.V. and V.A.; writing – review and editing were contributed by A.V., I.M., V.A., S.I.L., and P.M.; and supervision was provided by S.I.L. and P.M.

  3. Conflict of interest: Authors state no conflict of interest.

  4. Data availability statement: The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

References

[1] Kanjevac M, Zlatić N, Bojović B, Stanković M. Pharmaceutical and biological properties of Stachys species: A review. Braz J Pharm Sci. 2022;58:e20211.10.1590/s2175-97902022e20211Search in Google Scholar

[2] Tomou EM, Barda C, Skaltsa H. Genus stachys: A review of traditional uses, phytochemistry and bioactivity. Medicines. 2020 Sep;7(10):63.10.3390/medicines7100063Search in Google Scholar PubMed PubMed Central

[3] Benedec D, Oniga I, Hanganu D, Tiperciuc B, Nistor A, Vlase AM, et al. Stachys species: Comparative evaluation of phenolic profile and antimicrobial and antioxidant potential. Antibiotics. 2023 Nov;12(11):1644.10.3390/antibiotics12111644Search in Google Scholar PubMed PubMed Central

[4] Bahadori MB, Zengin G, Dinparast L, Eskandani M. The health benefits of three Hedgenettle herbal teas (Stachys byzantina, Stachys inflata, and Stachys lavandulifolia) - profiling phenolic and antioxidant activities. Eur J Integr Med. 2020 Jun;36:101134.10.1016/j.eujim.2020.101134Search in Google Scholar

[5] Slimani W, Maioli M, Cruciani S, Zerizer S, Santaniello S, Kabouche Z, et al. Antioxidant, anti-inflammatory and anti-proliferative properties of Stachys circinata on HepG2 and MCF7 cells. Plants. 2023 Jun;12(12):2272.10.3390/plants12122272Search in Google Scholar PubMed PubMed Central

[6] Mansourian M, Mirzaei A, Azarmehr N, Vakilpour H, Kokhdan EP, Doustimotlagh AH. Hepatoprotective and antioxidant activity of hydroalcoholic extract of Stachys pilifera. Benth on acetaminophen-induced liver toxicity in male rats. Heliyon. 2019 Dec;5(12):e03029.10.1016/j.heliyon.2019.e03029Search in Google Scholar PubMed PubMed Central

[7] Mantovska DI, Zhiponova MK, Georgiev MI, Alipieva K, Tsacheva I, Simova S, et al. Biological activity and NMR-fingerprinting of balkan endemic species Stachys thracica Davidov. Metabolites. 2022 Mar;12(3):251.10.3390/metabo12030251Search in Google Scholar PubMed PubMed Central

[8] Anagnostou M, Tomou E, Goya‐Jorge E, Chatzopoulou P, Giner RM, Skaltsa H. Phytochemical study of Stachys iva Griseb. and in vitro evaluation of AhR transcriptional activity. Chem Biodivers. 2024 Nov;21(11):e202400457.10.1002/cbdv.202400457Search in Google Scholar PubMed

[9] Farjam MH. Biological activity of the n-butanolic extract of Stachys pilifera. Afr J Microbiol Res. 2011 Nov;5(28):5115–9.10.5897/AJMR11.1066Search in Google Scholar

[10] Stegăruș DI, Lengyel E, Apostolescu GF, Botoran OR, Tanase C. Phytochemical analysis and biological activity of three Stachys species (Lamiaceae) from Romania. Plants. 2021 Dec;10(12):2710.10.3390/plants10122710Search in Google Scholar PubMed PubMed Central

[11] Napolitano A, Di Napoli M, Castagliuolo G, Badalamenti N, Cicio A, Bruno M, et al. The chemical composition of the aerial parts of Stachys spreitzenhoferi (Lamiaceae) growing in Kythira Island (Greece), and their antioxidant, antimicrobial, and antiproliferative properties. Phytochemistry. 2022 Nov;203:113373.10.1016/j.phytochem.2022.113373Search in Google Scholar PubMed

[12] Jaradat N, Hawash M, Al-Maharik N, Qadi M, Issa L, Sobuh S, et al. Characterization of volatile compounds and evaluation of antibacterial, antifungal, and cytotoxic properties of Stachys palaestina from Palestine. Arab J Sci Eng. 2024 Jul. [cited 2024 Dec 6] https://link.springer.com/10.1007/s13369-024-09306-w.10.1007/s13369-024-09306-wSearch in Google Scholar

[13] Pashova S, Karcheva-Bahchevanska D, Ivanov K, Ivanova S. Genus Stachys – phytochemistry, traditional medicinal uses, and future perspectives. Molecules. 2024 Nov;29(22):5345.10.3390/molecules29225345Search in Google Scholar PubMed PubMed Central

[14] Jassbi AR, Miri R, Asadollahi M, Javanmardi N, Firuzi O. Cytotoxic, antioxidant and antimicrobial effects of nine species of woundwort (Stachys) plants. Pharm Biol. 2014 Jan;52(1):62–7.10.3109/13880209.2013.810650Search in Google Scholar PubMed

[15] Duke JA, Bogenshcutz-Godwin MJ, duCellier J, Duke PAK. Handbook of medicinal herbs. 2nd edn. Boca Raton (FL): CRC Press; 2002.10.1201/9781420040463Search in Google Scholar

[16] Modarresi M, Hosseinzadeh L, Nematy N, Siavash-Haghighi ZM, Ghanbari K. Acute and subchronic toxicological evaluation of Stachys lavandulifolia aqueous extract in Wistar rats. Res Pharm Sci. 2014;9(3):165–72.Search in Google Scholar

[17] Aljerf L, Aljerf N. Food products quality and nutrition in relation to public. Balancing health and disease: Food safety & hygiene promotion. Prog Nutr. 2023 Mar;25(1):e2023024.Search in Google Scholar

[18] Pandey KB, Rizvi SI. Plant polyphenols as dietary antioxidants in human health and disease. Oxid Med Cell Longev. 2009 Jan;2(5):270–8.10.4161/oxim.2.5.9498Search in Google Scholar PubMed PubMed Central

[19] Onydinma UP, Aljerf L, Obike A, Onah OE, Caleb NJ. Evaluation of physicochemical characteristics and health risk of polycyclic aromatic hydrocarbons in borehole waters around automobile workshops in Southeastern Nigeria. Groundw Sustain Dev. 2021 Aug;14:100615.10.1016/j.gsd.2021.100615Search in Google Scholar

[20] Di Lorenzo C, Colombo F, Biella S, Stockley C, Restani P. Polyphenols and human health: The role of bioavailability. Nutrients. 2021 Jan;13(1):273.10.3390/nu13010273Search in Google Scholar PubMed PubMed Central

[21] Xu Z, Chenli Z. Inflammation. In: Chen K, Liang L, Li M, Pan Y, editors. Textbook of pathologic anatomy. Singapore: Springer Nature Singapore; 2024. p. 75–105. [cited 2024 Dec 5]. https://link.springer.com/10.1007/978-981-99-8445-9_4.10.1007/978-981-99-8445-9_4Search in Google Scholar

[22] Aljerf L, Williams M, Ajong AB, Onydinma UP, Dehmchi F, Pham VT, et al. Comparative study of the biochemical response behavior of some highly toxic minerals on selenosis in rats. Rev Chim. 2021 May;72(2):9–18.10.37358/RC.21.2.8415Search in Google Scholar

[23] Bai R, Guo J, Ye XY, Xie Y, Xie T. Oxidative stress: The core pathogenesis and mechanism of Alzheimer’s disease. Ageing Res Rev. 2022 May;77:101619.10.1016/j.arr.2022.101619Search in Google Scholar PubMed

[24] Hayes JD, Dinkova-Kostova AT, Tew KD. Oxidative stress in cancer. Cancer Cell. 2020 Aug;38(2):167–97.10.1016/j.ccell.2020.06.001Search in Google Scholar PubMed PubMed Central

[25] Jomova K, Raptova R, Alomar SY, Alwasel SH, Nepovimova E, Kuca K, et al. Reactive oxygen species, toxicity, oxidative stress, and antioxidants: Chronic diseases and aging. Arch Toxicol. 2023 Oct;97(10):2499–574.10.1007/s00204-023-03562-9Search in Google Scholar PubMed PubMed Central

[26] Rana A, Samtiya M, Dhewa T, Mishra V, Aluko RE. Health benefits of polyphenols: A concise review. J Food Biochem. 2022 Oct;46(10):e14264.10.1111/jfbc.14264Search in Google Scholar PubMed

[27] Pritsas A, Tomou EM, Tsitsigianni E, Papaemmanouil CD, Diamantis DA, Chatzopoulou P, et al. Valorisation of stachysetin from cultivated Stachys iva Griseb. as anti-diabetic agent: A multi-spectroscopic and molecular docking approach. J Biomol Struct Dyn. 2021 Nov;39(17):6452–66.10.1080/07391102.2020.1799864Search in Google Scholar PubMed

[28] Kokkini S, Karousou R, Hanlidou E. HERBS | Herbs of the labiatae. In: Caballero B, editor. Encyclopedia of food sciences and nutrition. 2nd ed. London: Academic Press; 2003. p. 3082–90.10.1016/B0-12-227055-X/00593-9Search in Google Scholar

[29] Aljerf L. High-efficiency extraction of bromocresol purple dye and heavy metals as chromium from industrial effluent by adsorption onto a modified surface of zeolite: Kinetics and equilibrium study. J Environ Manag. 2018 Nov;225:120–32.10.1016/j.jenvman.2018.07.048Search in Google Scholar PubMed

[30] Chatzimitakos T, Athanasiadis V, Makrygiannis I, Kalompatsios D, Bozinou E, Lalas SI. An investigation into Crithmum maritimum L. leaves as a source of antioxidant polyphenols. Compounds. 2023 Dec;3(4):532–51.10.3390/compounds3040038Search in Google Scholar

[31] Shehata E, Grigorakis S, Loupassaki S, Makris DP. Extraction optimisation using water/glycerol for the efficient recovery of polyphenolic antioxidants from two Artemisia species. Sep Purif Technol. 2015 Jul;149:462–9.10.1016/j.seppur.2015.06.017Search in Google Scholar

[32] Chatzimitakos T, Athanasiadis V, Kotsou K, Bozinou E, Lalas SI. Response surface optimization for the enhancement of the extraction of bioactive compounds from Citrus limon peel. Antioxidants. 2023 Aug;12(8):1605.10.3390/antiox12081605Search in Google Scholar PubMed PubMed Central

[33] Kasouni AI, Chatzimitakos TG, Stalikas CD, Trangas T, Papoudou-Bai A, Troganis AN. The unexplored wound healing activity of Urtica dioica L. extract: An in vitro and in vivo study. Molecules. 2021 Oct;26(20):6248.10.3390/molecules26206248Search in Google Scholar PubMed PubMed Central

[34] Dirar AI, Alsaadi DHM, Wada M, Mohamed MA, Watanabe T, Devkota HP. Effects of extraction solvents on total phenolic and flavonoid contents and biological activities of extracts from Sudanese medicinal plants. South Afr J Botany. 2019 Jan;120:261–7.10.1016/j.sajb.2018.07.003Search in Google Scholar

[35] Thong-on W, Pathomwichaiwat T, Boonsith S, Koo-amornpattana W, Prathanturarug S. Green extraction optimization of triterpenoid glycoside-enriched extract from Centella asiatica (L.) Urban using response surface methodology (RSM). Sci Rep. 2021 Nov;11(1):22026.10.1038/s41598-021-01602-xSearch in Google Scholar PubMed PubMed Central

[36] Osorio-Tobón JF. Recent advances and comparisons of conventional and alternative extraction techniques of phenolic compounds. J Food Sci Technol. 2020 Dec;57(12):4299–315.10.1007/s13197-020-04433-2Search in Google Scholar PubMed PubMed Central

[37] Luo C, Zou L, Sun H, Peng J, Gao C, Bao L, et al. A review of the anti-inflammatory effects of rosmarinic acid on inflammatory diseases. Front Pharmacol. 2020 Feb;11:153.10.3389/fphar.2020.00153Search in Google Scholar PubMed PubMed Central

[38] Guan H, Luo W, Bao B, Cao Y, Cheng F, Yu S, et al. A comprehensive review of rosmarinic acid: From phytochemistry to pharmacology and its new insight. Molecules. 2022 May;27(10):3292.10.3390/molecules27103292Search in Google Scholar PubMed PubMed Central

[39] Dahchour A. Anxiolytic and antidepressive potentials of rosmarinic acid: A review with a focus on antioxidant and anti-inflammatory effects. Pharmacol Res. 2022 Oct;184:106421.10.1016/j.phrs.2022.106421Search in Google Scholar PubMed

[40] Kernou ON, Azzouz Z, Madani K, Rijo P. Application of rosmarinic acid with its derivatives in the treatment of microbial pathogens. Molecules. 2023 May;28(10):4243.10.3390/molecules28104243Search in Google Scholar PubMed PubMed Central

[41] Miao M, Xiang L. Pharmacological action and potential targets of chlorogenic acid. In Advances in pharmacology. London: Academic Press; 2020. p. 71–88.10.1016/bs.apha.2019.12.002Search in Google Scholar PubMed

[42] Nguyen V, Taine EG, Meng D, Cui T, Tan W. Chlorogenic acid: A systematic review on the biological functions, mechanistic actions, and therapeutic potentials. Nutrients. 2024 Mar;16(7):924.10.3390/nu16070924Search in Google Scholar PubMed PubMed Central

[43] Huang J, Xie M, He L, Song X, Cao T. Chlorogenic acid: A review on its mechanisms of anti-inflammation, disease treatment, and related delivery systems. Front Pharmacol. 2023 Sep;14:1218015.10.3389/fphar.2023.1218015Search in Google Scholar PubMed PubMed Central

[44] Park SY, Jin ML, Yi EH, Kim Y, Park G. Neochlorogenic acid inhibits against LPS-activated inflammatory responses through up-regulation of Nrf2/HO-1 and involving AMPK pathway. Environ Toxicol Pharmacology. 2018 Sep;62:1–10.10.1016/j.etap.2018.06.001Search in Google Scholar PubMed

[45] Gao XH, Zhang SD, Wang LT, Yu L, Zhao XL, Ni HY, et al. Anti-inflammatory effects of neochlorogenic acid extract from mulberry leaf (Morus alba L.) against LPS-stimulated inflammatory response through mediating the AMPK/Nrf2 signaling pathway in A549 cells. Molecules. 2020 Mar;25(6):1385.10.3390/molecules25061385Search in Google Scholar PubMed PubMed Central

[46] Zhao Y, Wang S, Pan J, Ma K. Verbascoside: A neuroprotective phenylethanoid glycosides with anti-depressive properties. Phytomedicine. 2023 Nov;120:155027.10.1016/j.phymed.2023.155027Search in Google Scholar PubMed

[47] Pongkitwitoon B, Putalun W, Triwitayakorn K, Kitisripanya T, Kanchanapoom T, Boonsnongcheep P. Anti-inflammatory activity of verbascoside- and isoverbascoside-rich Lamiales medicinal plants. Heliyon. 2024 Jan;10(1):e23644.10.1016/j.heliyon.2023.e23644Search in Google Scholar PubMed PubMed Central

[48] Lei P, Lü J, Yao T, Zhang P, Chai X, Wang Y, et al. Verbascoside exerts an anti-atherosclerotic effect by regulating liver glycerophospholipid metabolism. Food Sci Hum Wellness. 2023 Nov;12(6):2314–23.10.1016/j.fshw.2023.03.035Search in Google Scholar

[49] Kumar S, Swamy N, Tuli HS, Rani S, Garg A, Mishra D, et al. Myricetin: A potential plant-derived anticancer bioactive compound – an updated overview. Naunyn-Schmiedeberg’s Arch Pharmacol. 2023 Oct;396(10):2179–96.10.1007/s00210-023-02479-5Search in Google Scholar PubMed

[50] Agraharam G, Girigoswami A, Girigoswami K. Myricetin: A multifunctional flavonol in biomedicine. Curr Pharmacol Rep. 2022 Feb;8(1):48–61.10.1007/s40495-021-00269-2Search in Google Scholar PubMed PubMed Central

[51] Imani A, Maleki N, Bohlouli S, Kouhsoltani M, Sharifi S, Maleki Dizaj S. Molecular mechanisms of anticancer effect of rutin. Phytother Res. 2021 May;35(5):2500–13.10.1002/ptr.6977Search in Google Scholar PubMed

[52] Farha AK, Gan RY, Li HB, Wu DT, Atanasov AG, Gul K, et al. The anticancer potential of the dietary polyphenol rutin: Current status, challenges, and perspectives. Crit Rev Food Sci Nutr. 2022 Jan;62(3):832–59.10.1080/10408398.2020.1829541Search in Google Scholar PubMed

[53] Choi SS, Park HR, Lee KA. A comparative study of rutin and rutin glycoside: Antioxidant activity, anti-inflammatory effect, effect on platelet aggregation and blood coagulation. Antioxidants. 2021 Oct;10(11):1696.10.3390/antiox10111696Search in Google Scholar PubMed PubMed Central

[54] Taheri Y, Suleria HAR, Martins N, Sytar O, Beyatli A, Yeskaliyeva B, et al. Myricetin bioactive effects: Moving from preclinical evidence to potential clinical applications. BMC Complement Med Ther. 2020 Dec;20(1):241.10.1186/s12906-020-03033-zSearch in Google Scholar PubMed PubMed Central

[55] Imran M, Saeed F, Hussain G, Imran A, Mehmood Z, Gondal TA, et al. Myricetin: A comprehensive review on its biological potentials. Food Sci & Nutr. 2021 Oct;9(10):5854–68.10.1002/fsn3.2513Search in Google Scholar PubMed PubMed Central

[56] Mitra S, Lami MS, Uddin TM, Das R, Islam F, Anjum J, et al. Prospective multifunctional roles and pharmacological potential of dietary flavonoid narirutin. Biomed Pharmacother. 2022 Jun;150:112932.10.1016/j.biopha.2022.112932Search in Google Scholar PubMed

[57] Pandey P, Khan F, Ramniwas S, Saeed M, Ahmad I. A mechanistic review of the pharmacological potential of narirutin: A dietary flavonoid. Naunyn-Schmiedeberg’s Arch Pharmacol. 2024 Aug;397(8):5449–61.10.1007/s00210-024-03022-wSearch in Google Scholar PubMed

[58] Ri MH, Li MY, Xing Y, Zuo HX, Li G, Li C, et al. Narirutin exerts anti‐inflammatory activity by inhibiting NLRP3 inflammasome activation in macrophages. Phytotherapy Res. 2023 Apr;37(4):1293–308.10.1002/ptr.7686Search in Google Scholar PubMed

[59] Qurtam AA, Mechchate H, Es-safi I, Al-zharani M, Nasr FA, Noman OM, et al. Citrus flavanone narirutin, in vitro and in silico mechanistic antidiabetic potential. Pharmaceutics. 2021 Oct;13(11):1818.10.3390/pharmaceutics13111818Search in Google Scholar PubMed PubMed Central

Received: 2024-10-21
Revised: 2024-12-08
Accepted: 2024-12-27
Published Online: 2025-02-19

© 2025 the author(s), published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

Articles in the same Issue

  1. Biomedical Sciences
  2. Mechanism of triptolide regulating proliferation and apoptosis of hepatoma cells by inhibiting JAK/STAT pathway
  3. Maslinic acid improves mitochondrial function and inhibits oxidative stress and autophagy in human gastric smooth muscle cells
  4. Comparative analysis of inflammatory biomarkers for the diagnosis of neonatal sepsis: IL-6, IL-8, SAA, CRP, and PCT
  5. Post-pandemic insights on COVID-19 and premature ovarian insufficiency
  6. Proteome differences of dental stem cells between permanent and deciduous teeth by data-independent acquisition proteomics
  7. Optimizing a modified cetyltrimethylammonium bromide protocol for fungal DNA extraction: Insights from multilocus gene amplification
  8. Preliminary analysis of the role of small hepatitis B surface proteins mutations in the pathogenesis of occult hepatitis B infection via the endoplasmic reticulum stress-induced UPR-ERAD pathway
  9. Efficacy of alginate-coated gold nanoparticles against antibiotics-resistant Staphylococcus and Streptococcus pathogens of acne origins
  10. Battling COVID-19 leveraging nanobiotechnology: Gold and silver nanoparticle–B-escin conjugates as SARS-CoV-2 inhibitors
  11. Neurodegenerative diseases and neuroinflammation-induced apoptosis
  12. Impact of fracture fixation surgery on cognitive function and the gut microbiota in mice with a history of stroke
  13. COLEC10: A potential tumor suppressor and prognostic biomarker in hepatocellular carcinoma through modulation of EMT and PI3K-AKT pathways
  14. High-temperature requirement serine protease A2 inhibitor UCF-101 ameliorates damaged neurons in traumatic brain-injured rats by the AMPK/NF-κB pathway
  15. SIK1 inhibits IL-1β-stimulated cartilage apoptosis and inflammation in vitro through the CRTC2/CREB1 signaling
  16. Rutin–chitooligosaccharide complex: Comprehensive evaluation of its anti-inflammatory and analgesic properties in vitro and in vivo
  17. Knockdown of Aurora kinase B alleviates high glucose-triggered trophoblast cells damage and inflammation during gestational diabetes
  18. Calcium-sensing receptors promoted Homer1 expression and osteogenic differentiation in bone marrow mesenchymal stem cells
  19. ABI3BP can inhibit the proliferation, invasion, and epithelial–mesenchymal transition of non-small-cell lung cancer cells
  20. Changes in blood glucose and metabolism in hyperuricemia mice
  21. Rapid detection of the GJB2 c.235delC mutation based on CRISPR-Cas13a combined with lateral flow dipstick
  22. IL-11 promotes Ang II-induced autophagy inhibition and mitochondrial dysfunction in atrial fibroblasts
  23. Short-chain fatty acid attenuates intestinal inflammation by regulation of gut microbial composition in antibiotic-associated diarrhea
  24. Application of metagenomic next-generation sequencing in the diagnosis of pathogens in patients with diabetes complicated by community-acquired pneumonia
  25. NAT10 promotes radiotherapy resistance in non-small cell lung cancer by regulating KPNB1-mediated PD-L1 nuclear translocation
  26. Phytol-mixed micelles alleviate dexamethasone-induced osteoporosis in zebrafish: Activation of the MMP3–OPN–MAPK pathway-mediating bone remodeling
  27. Association between TGF-β1 and β-catenin expression in the vaginal wall of patients with pelvic organ prolapse
  28. Primary pleomorphic liposarcoma involving bilateral ovaries: Case report and literature review
  29. Effects of de novo donor-specific Class I and II antibodies on graft outcomes after liver transplantation: A pilot cohort study
  30. Sleep architecture in Alzheimer’s disease continuum: The deep sleep question
  31. Ephedra fragilis plant extract: A groundbreaking corrosion inhibitor for mild steel in acidic environments – electrochemical, EDX, DFT, and Monte Carlo studies
  32. Langerhans cell histiocytosis in an adult patient with upper jaw and pulmonary involvement: A case report
  33. Inhibition of mast cell activation by Jaranol-targeted Pirin ameliorates allergic responses in mouse allergic rhinitis
  34. Aeromonas veronii-induced septic arthritis of the hip in a child with acute lymphoblastic leukemia
  35. Clusterin activates the heat shock response via the PI3K/Akt pathway to protect cardiomyocytes from high-temperature-induced apoptosis
  36. Research progress on fecal microbiota transplantation in tumor prevention and treatment
  37. Low-pressure exposure influences the development of HAPE
  38. Stigmasterol alleviates endplate chondrocyte degeneration through inducing mitophagy by enhancing PINK1 mRNA acetylation via the ESR1/NAT10 axis
  39. AKAP12, mediated by transcription factor 21, inhibits cell proliferation, metastasis, and glycolysis in lung squamous cell carcinoma
  40. Association between PAX9 or MSX1 gene polymorphism and tooth agenesis risk: A meta-analysis
  41. A case of bloodstream infection caused by Neisseria gonorrhoeae
  42. Case of nasopharyngeal tuberculosis complicated with cervical lymph node and pulmonary tuberculosis
  43. p-Cymene inhibits pro-fibrotic and inflammatory mediators to prevent hepatic dysfunction
  44. GFPT2 promotes paclitaxel resistance in epithelial ovarian cancer cells via activating NF-κB signaling pathway
  45. Transfer RNA-derived fragment tRF-36 modulates varicose vein progression via human vascular smooth muscle cell Notch signaling
  46. RTA-408 attenuates the hepatic ischemia reperfusion injury in mice possibly by activating the Nrf2/HO-1 signaling pathway
  47. Decreased serum TIMP4 levels in patients with rheumatoid arthritis
  48. Sirt1 protects lupus nephritis by inhibiting the NLRP3 signaling pathway in human glomerular mesangial cells
  49. Sodium butyrate aids brain injury repair in neonatal rats
  50. Interaction of MTHFR polymorphism with PAX1 methylation in cervical cancer
  51. Convallatoxin inhibits proliferation and angiogenesis of glioma cells via regulating JAK/STAT3 pathway
  52. The effect of the PKR inhibitor, 2-aminopurine, on the replication of influenza A virus, and segment 8 mRNA splicing
  53. Effects of Ire1 gene on virulence and pathogenicity of Candida albicans
  54. Small cell lung cancer with small intestinal metastasis: Case report and literature review
  55. GRB14: A prognostic biomarker driving tumor progression in gastric cancer through the PI3K/AKT signaling pathway by interacting with COBLL1
  56. 15-Lipoxygenase-2 deficiency induces foam cell formation that can be restored by salidroside through the inhibition of arachidonic acid effects
  57. FTO alleviated the diabetic nephropathy progression by regulating the N6-methyladenosine levels of DACT1
  58. Clinical relevance of inflammatory markers in the evaluation of severity of ulcerative colitis: A retrospective study
  59. Zinc valproic acid complex promotes osteoblast differentiation and exhibits anti-osteoporotic potential
  60. Primary pulmonary synovial sarcoma in the bronchial cavity: A case report
  61. Metagenomic next-generation sequencing of alveolar lavage fluid improves the detection of pulmonary infection
  62. Uterine tumor resembling ovarian sex cord tumor with extensive rhabdoid differentiation: A case report
  63. Genomic analysis of a novel ST11(PR34365) Clostridioides difficile strain isolated from the human fecal of a CDI patient in Guizhou, China
  64. Effects of tiered cardiac rehabilitation on CRP, TNF-α, and physical endurance in older adults with coronary heart disease
  65. Changes in T-lymphocyte subpopulations in patients with colorectal cancer before and after acupoint catgut embedding acupuncture observation
  66. Modulating the tumor microenvironment: The role of traditional Chinese medicine in improving lung cancer treatment
  67. Alterations of metabolites related to microbiota–gut–brain axis in plasma of colon cancer, esophageal cancer, stomach cancer, and lung cancer patients
  68. Research on individualized drug sensitivity detection technology based on bio-3D printing technology for precision treatment of gastrointestinal stromal tumors
  69. CEBPB promotes ulcerative colitis-associated colorectal cancer by stimulating tumor growth and activating the NF-κB/STAT3 signaling pathway
  70. Oncolytic bacteria: A revolutionary approach to cancer therapy
  71. A de novo meningioma with rapid growth: A possible malignancy imposter?
  72. Diagnosis of secondary tuberculosis infection in an asymptomatic elderly with cancer using next-generation sequencing: Case report
  73. Hesperidin and its zinc(ii) complex enhance osteoblast differentiation and bone formation: In vitro and in vivo evaluations
  74. Research progress on the regulation of autophagy in cardiovascular diseases by chemokines
  75. Anti-arthritic, immunomodulatory, and inflammatory regulation by the benzimidazole derivative BMZ-AD: Insights from an FCA-induced rat model
  76. Immunoassay for pyruvate kinase M1/2 as an Alzheimer’s biomarker in CSF
  77. The role of HDAC11 in age-related hearing loss: Mechanisms and therapeutic implications
  78. Evaluation and application analysis of animal models of PIPNP based on data mining
  79. Therapeutic approaches for liver fibrosis/cirrhosis by targeting pyroptosis
  80. Fabrication of zinc oxide nanoparticles using Ruellia tuberosa leaf extract induces apoptosis through P53 and STAT3 signalling pathways in prostate cancer cells
  81. Haplo-hematopoietic stem cell transplantation and immunoradiotherapy for severe aplastic anemia complicated with nasopharyngeal carcinoma: A case report
  82. Modulation of the KEAP1-NRF2 pathway by Erianin: A novel approach to reduce psoriasiform inflammation and inflammatory signaling
  83. The expression of epidermal growth factor receptor 2 and its relationship with tumor-infiltrating lymphocytes and clinical pathological features in breast cancer patients
  84. Innovations in MALDI-TOF Mass Spectrometry: Bridging modern diagnostics and historical insights
  85. BAP1 complexes with YY1 and RBBP7 and its downstream targets in ccRCC cells
  86. Hypereosinophilic syndrome with elevated IgG4 and T-cell clonality: A report of two cases
  87. Electroacupuncture alleviates sciatic nerve injury in sciatica rats by regulating BDNF and NGF levels, myelin sheath degradation, and autophagy
  88. Polydatin prevents cholesterol gallstone formation by regulating cholesterol metabolism via PPAR-γ signaling
  89. RNF144A and RNF144B: Important molecules for health
  90. Analysis of the detection rate and related factors of thyroid nodules in the healthy population
  91. Artesunate inhibits hepatocellular carcinoma cell migration and invasion through OGA-mediated O-GlcNAcylation of ZEB1
  92. Endovascular management of post-pancreatectomy hemorrhage caused by a hepatic artery pseudoaneurysm: Case report and review of the literature
  93. Efficacy and safety of anti-PD-1/PD-L1 antibodies in patients with relapsed refractory diffuse large B-cell lymphoma: A meta-analysis
  94. SATB2 promotes humeral fracture healing in rats by activating the PI3K/AKT pathway
  95. Overexpression of the ferroptosis-related gene, NFS1, corresponds to gastric cancer growth and tumor immune infiltration
  96. Understanding risk factors and prognosis in diabetic foot ulcers
  97. Atractylenolide I alleviates the experimental allergic response in mice by suppressing TLR4/NF-kB/NLRP3 signalling
  98. FBXO31 inhibits the stemness characteristics of CD147 (+) melanoma stem cells
  99. Immune molecule diagnostics in colorectal cancer: CCL2 and CXCL11
  100. Inhibiting CXCR6 promotes senescence of activated hepatic stellate cells with limited proinflammatory SASP to attenuate hepatic fibrosis
  101. Cadmium toxicity, health risk and its remediation using low-cost biochar adsorbents
  102. Pulmonary cryptococcosis with headache as the first presentation: A case report
  103. Solitary pulmonary metastasis with cystic airspaces in colon cancer: A rare case report
  104. RUNX1 promotes denervation-induced muscle atrophy by activating the JUNB/NF-κB pathway and driving M1 macrophage polarization
  105. Morphometric analysis and immunobiological investigation of Indigofera oblongifolia on the infected lung with Plasmodium chabaudi
  106. The NuA4/TIP60 histone-modifying complex and Hr78 modulate the Lobe2 mutant eye phenotype
  107. Experimental study on salmon demineralized bone matrix loaded with recombinant human bone morphogenetic protein-2: In vitro and in vivo study
  108. A case of IgA nephropathy treated with a combination of telitacicept and half-dose glucocorticoids
  109. Analgesic and toxicological evaluation of cannabidiol-rich Moroccan Cannabis sativa L. (Khardala variety) extract: Evidence from an in vivo and in silico study
  110. Wound healing and signaling pathways
  111. Combination of immunotherapy and whole-brain radiotherapy on prognosis of patients with multiple brain metastases: A retrospective cohort study
  112. To explore the relationship between endometrial hyperemia and polycystic ovary syndrome
  113. Research progress on the impact of curcumin on immune responses in breast cancer
  114. Biogenic Cu/Ni nanotherapeutics from Descurainia sophia (L.) Webb ex Prantl seeds for the treatment of lung cancer
  115. Dapagliflozin attenuates atrial fibrosis via the HMGB1/RAGE pathway in atrial fibrillation rats
  116. Ecology and Environmental Science
  117. Optimization and comparative study of Bacillus consortia for cellulolytic potential and cellulase enzyme activity
  118. The complete mitochondrial genome analysis of Haemaphysalis hystricis Supino, 1897 (Ixodida: Ixodidae) and its phylogenetic implications
  119. Epidemiological characteristics and risk factors analysis of multidrug-resistant tuberculosis among tuberculosis population in Huzhou City, Eastern China
  120. Indices of human impacts on landscapes: How do they reflect the proportions of natural habitats?
  121. Genetic analysis of the Siberian flying squirrel population in the northern Changbai Mountains, Northeast China: Insights into population status and conservation
  122. Diversity and environmental drivers of Suillus communities in Pinus sylvestris var. mongolica forests of Inner Mongolia
  123. Agriculture
  124. Integrated analysis of transcriptome, sRNAome, and degradome involved in the drought-response of maize Zhengdan958
  125. Variation in flower frost tolerance among seven apple cultivars and transcriptome response patterns in two contrastingly frost-tolerant selected cultivars
  126. Heritability of durable resistance to stripe rust in bread wheat (Triticum aestivum L.)
  127. Animal Science
  128. Effect of sex ratio on the life history traits of an important invasive species, Spodoptera frugiperda
  129. Plant Sciences
  130. Hairpin in a haystack: In silico identification and characterization of plant-conserved microRNA in Rafflesiaceae
  131. Widely targeted metabolomics of different tissues in Rubus corchorifolius
  132. The complete chloroplast genome of Gerbera piloselloides (L.) Cass., 1820 (Carduoideae, Asteraceae) and its phylogenetic analysis
  133. Field trial to correlate mineral solubilization activity of Pseudomonas aeruginosa and biochemical content of groundnut plants
  134. Correlation analysis between semen routine parameters and sperm DNA fragmentation index in patients with semen non-liquefaction: A retrospective study
  135. Plasticity of the anatomical traits of Rhododendron L. (Ericaceae) leaves and its implications in adaptation to the plateau environment
  136. Effects of Piriformospora indica and arbuscular mycorrhizal fungus on growth and physiology of Moringa oleifera under low-temperature stress
  137. Effects of different sources of potassium fertiliser on yield, fruit quality and nutrient absorption in “Harward” kiwifruit (Actinidia deliciosa)
  138. Comparative efficiency and residue levels of spraying programs against powdery mildew in grape varieties
  139. The DREB7 transcription factor enhances salt tolerance in soybean plants under salt stress
  140. Food Science
  141. Phytochemical analysis of Stachys iva: Discovering the optimal extract conditions and its bioactive compounds
  142. Review on role of honey in disease prevention and treatment through modulation of biological activities
  143. Computational analysis of polymorphic residues in maltose and maltotriose transporters of a wild Saccharomyces cerevisiae strain
  144. Optimization of phenolic compound extraction from Tunisian squash by-products: A sustainable approach for antioxidant and antibacterial applications
  145. Liupao tea aqueous extract alleviates dextran sulfate sodium-induced ulcerative colitis in rats by modulating the gut microbiota
  146. Toxicological qualities and detoxification trends of fruit by-products for valorization: A review
  147. Polyphenolic spectrum of cornelian cherry fruits and their health-promoting effect
  148. Optimizing the encapsulation of the refined extract of squash peels for functional food applications: A sustainable approach to reduce food waste
  149. Advancements in curcuminoid formulations: An update on bioavailability enhancement strategies curcuminoid bioavailability and formulations
  150. Impact of saline sprouting on antioxidant properties and bioactive compounds in chia seeds
  151. The dilemma of food genetics and improvement
  152. Bioengineering and Biotechnology
  153. Impact of hyaluronic acid-modified hafnium metalorganic frameworks containing rhynchophylline on Alzheimer’s disease
  154. Emerging patterns in nanoparticle-based therapeutic approaches for rheumatoid arthritis: A comprehensive bibliometric and visual analysis spanning two decades
  155. Application of CRISPR/Cas gene editing for infectious disease control in poultry
  156. Preparation of hafnium nitride-coated titanium implants by magnetron sputtering technology and evaluation of their antibacterial properties and biocompatibility
  157. Preparation and characterization of lemongrass oil nanoemulsion: Antimicrobial, antibiofilm, antioxidant, and anticancer activities
  158. Corrigendum
  159. Corrigendum to “Utilization of convolutional neural networks to analyze microscopic images for high-throughput screening of mesenchymal stem cells”
https://doi.org/10.1515/biol-2022-1053
Keywords for this article
Stachys; optimization; polyphenols; antioxidant; anti-inflammatory
Creative Commons
BY 4.0

Articles in the same Issue

  1. Biomedical Sciences
  2. Mechanism of triptolide regulating proliferation and apoptosis of hepatoma cells by inhibiting JAK/STAT pathway
  3. Maslinic acid improves mitochondrial function and inhibits oxidative stress and autophagy in human gastric smooth muscle cells
  4. Comparative analysis of inflammatory biomarkers for the diagnosis of neonatal sepsis: IL-6, IL-8, SAA, CRP, and PCT
  5. Post-pandemic insights on COVID-19 and premature ovarian insufficiency
  6. Proteome differences of dental stem cells between permanent and deciduous teeth by data-independent acquisition proteomics
  7. Optimizing a modified cetyltrimethylammonium bromide protocol for fungal DNA extraction: Insights from multilocus gene amplification
  8. Preliminary analysis of the role of small hepatitis B surface proteins mutations in the pathogenesis of occult hepatitis B infection via the endoplasmic reticulum stress-induced UPR-ERAD pathway
  9. Efficacy of alginate-coated gold nanoparticles against antibiotics-resistant Staphylococcus and Streptococcus pathogens of acne origins
  10. Battling COVID-19 leveraging nanobiotechnology: Gold and silver nanoparticle–B-escin conjugates as SARS-CoV-2 inhibitors
  11. Neurodegenerative diseases and neuroinflammation-induced apoptosis
  12. Impact of fracture fixation surgery on cognitive function and the gut microbiota in mice with a history of stroke
  13. COLEC10: A potential tumor suppressor and prognostic biomarker in hepatocellular carcinoma through modulation of EMT and PI3K-AKT pathways
  14. High-temperature requirement serine protease A2 inhibitor UCF-101 ameliorates damaged neurons in traumatic brain-injured rats by the AMPK/NF-κB pathway
  15. SIK1 inhibits IL-1β-stimulated cartilage apoptosis and inflammation in vitro through the CRTC2/CREB1 signaling
  16. Rutin–chitooligosaccharide complex: Comprehensive evaluation of its anti-inflammatory and analgesic properties in vitro and in vivo
  17. Knockdown of Aurora kinase B alleviates high glucose-triggered trophoblast cells damage and inflammation during gestational diabetes
  18. Calcium-sensing receptors promoted Homer1 expression and osteogenic differentiation in bone marrow mesenchymal stem cells
  19. ABI3BP can inhibit the proliferation, invasion, and epithelial–mesenchymal transition of non-small-cell lung cancer cells
  20. Changes in blood glucose and metabolism in hyperuricemia mice
  21. Rapid detection of the GJB2 c.235delC mutation based on CRISPR-Cas13a combined with lateral flow dipstick
  22. IL-11 promotes Ang II-induced autophagy inhibition and mitochondrial dysfunction in atrial fibroblasts
  23. Short-chain fatty acid attenuates intestinal inflammation by regulation of gut microbial composition in antibiotic-associated diarrhea
  24. Application of metagenomic next-generation sequencing in the diagnosis of pathogens in patients with diabetes complicated by community-acquired pneumonia
  25. NAT10 promotes radiotherapy resistance in non-small cell lung cancer by regulating KPNB1-mediated PD-L1 nuclear translocation
  26. Phytol-mixed micelles alleviate dexamethasone-induced osteoporosis in zebrafish: Activation of the MMP3–OPN–MAPK pathway-mediating bone remodeling
  27. Association between TGF-β1 and β-catenin expression in the vaginal wall of patients with pelvic organ prolapse
  28. Primary pleomorphic liposarcoma involving bilateral ovaries: Case report and literature review
  29. Effects of de novo donor-specific Class I and II antibodies on graft outcomes after liver transplantation: A pilot cohort study
  30. Sleep architecture in Alzheimer’s disease continuum: The deep sleep question
  31. Ephedra fragilis plant extract: A groundbreaking corrosion inhibitor for mild steel in acidic environments – electrochemical, EDX, DFT, and Monte Carlo studies
  32. Langerhans cell histiocytosis in an adult patient with upper jaw and pulmonary involvement: A case report
  33. Inhibition of mast cell activation by Jaranol-targeted Pirin ameliorates allergic responses in mouse allergic rhinitis
  34. Aeromonas veronii-induced septic arthritis of the hip in a child with acute lymphoblastic leukemia
  35. Clusterin activates the heat shock response via the PI3K/Akt pathway to protect cardiomyocytes from high-temperature-induced apoptosis
  36. Research progress on fecal microbiota transplantation in tumor prevention and treatment
  37. Low-pressure exposure influences the development of HAPE
  38. Stigmasterol alleviates endplate chondrocyte degeneration through inducing mitophagy by enhancing PINK1 mRNA acetylation via the ESR1/NAT10 axis
  39. AKAP12, mediated by transcription factor 21, inhibits cell proliferation, metastasis, and glycolysis in lung squamous cell carcinoma
  40. Association between PAX9 or MSX1 gene polymorphism and tooth agenesis risk: A meta-analysis
  41. A case of bloodstream infection caused by Neisseria gonorrhoeae
  42. Case of nasopharyngeal tuberculosis complicated with cervical lymph node and pulmonary tuberculosis
  43. p-Cymene inhibits pro-fibrotic and inflammatory mediators to prevent hepatic dysfunction
  44. GFPT2 promotes paclitaxel resistance in epithelial ovarian cancer cells via activating NF-κB signaling pathway
  45. Transfer RNA-derived fragment tRF-36 modulates varicose vein progression via human vascular smooth muscle cell Notch signaling
  46. RTA-408 attenuates the hepatic ischemia reperfusion injury in mice possibly by activating the Nrf2/HO-1 signaling pathway
  47. Decreased serum TIMP4 levels in patients with rheumatoid arthritis
  48. Sirt1 protects lupus nephritis by inhibiting the NLRP3 signaling pathway in human glomerular mesangial cells
  49. Sodium butyrate aids brain injury repair in neonatal rats
  50. Interaction of MTHFR polymorphism with PAX1 methylation in cervical cancer
  51. Convallatoxin inhibits proliferation and angiogenesis of glioma cells via regulating JAK/STAT3 pathway
  52. The effect of the PKR inhibitor, 2-aminopurine, on the replication of influenza A virus, and segment 8 mRNA splicing
  53. Effects of Ire1 gene on virulence and pathogenicity of Candida albicans
  54. Small cell lung cancer with small intestinal metastasis: Case report and literature review
  55. GRB14: A prognostic biomarker driving tumor progression in gastric cancer through the PI3K/AKT signaling pathway by interacting with COBLL1
  56. 15-Lipoxygenase-2 deficiency induces foam cell formation that can be restored by salidroside through the inhibition of arachidonic acid effects
  57. FTO alleviated the diabetic nephropathy progression by regulating the N6-methyladenosine levels of DACT1
  58. Clinical relevance of inflammatory markers in the evaluation of severity of ulcerative colitis: A retrospective study
  59. Zinc valproic acid complex promotes osteoblast differentiation and exhibits anti-osteoporotic potential
  60. Primary pulmonary synovial sarcoma in the bronchial cavity: A case report
  61. Metagenomic next-generation sequencing of alveolar lavage fluid improves the detection of pulmonary infection
  62. Uterine tumor resembling ovarian sex cord tumor with extensive rhabdoid differentiation: A case report
  63. Genomic analysis of a novel ST11(PR34365) Clostridioides difficile strain isolated from the human fecal of a CDI patient in Guizhou, China
  64. Effects of tiered cardiac rehabilitation on CRP, TNF-α, and physical endurance in older adults with coronary heart disease
  65. Changes in T-lymphocyte subpopulations in patients with colorectal cancer before and after acupoint catgut embedding acupuncture observation
  66. Modulating the tumor microenvironment: The role of traditional Chinese medicine in improving lung cancer treatment
  67. Alterations of metabolites related to microbiota–gut–brain axis in plasma of colon cancer, esophageal cancer, stomach cancer, and lung cancer patients
  68. Research on individualized drug sensitivity detection technology based on bio-3D printing technology for precision treatment of gastrointestinal stromal tumors
  69. CEBPB promotes ulcerative colitis-associated colorectal cancer by stimulating tumor growth and activating the NF-κB/STAT3 signaling pathway
  70. Oncolytic bacteria: A revolutionary approach to cancer therapy
  71. A de novo meningioma with rapid growth: A possible malignancy imposter?
  72. Diagnosis of secondary tuberculosis infection in an asymptomatic elderly with cancer using next-generation sequencing: Case report
  73. Hesperidin and its zinc(ii) complex enhance osteoblast differentiation and bone formation: In vitro and in vivo evaluations
  74. Research progress on the regulation of autophagy in cardiovascular diseases by chemokines
  75. Anti-arthritic, immunomodulatory, and inflammatory regulation by the benzimidazole derivative BMZ-AD: Insights from an FCA-induced rat model
  76. Immunoassay for pyruvate kinase M1/2 as an Alzheimer’s biomarker in CSF
  77. The role of HDAC11 in age-related hearing loss: Mechanisms and therapeutic implications
  78. Evaluation and application analysis of animal models of PIPNP based on data mining
  79. Therapeutic approaches for liver fibrosis/cirrhosis by targeting pyroptosis
  80. Fabrication of zinc oxide nanoparticles using Ruellia tuberosa leaf extract induces apoptosis through P53 and STAT3 signalling pathways in prostate cancer cells
  81. Haplo-hematopoietic stem cell transplantation and immunoradiotherapy for severe aplastic anemia complicated with nasopharyngeal carcinoma: A case report
  82. Modulation of the KEAP1-NRF2 pathway by Erianin: A novel approach to reduce psoriasiform inflammation and inflammatory signaling
  83. The expression of epidermal growth factor receptor 2 and its relationship with tumor-infiltrating lymphocytes and clinical pathological features in breast cancer patients
  84. Innovations in MALDI-TOF Mass Spectrometry: Bridging modern diagnostics and historical insights
  85. BAP1 complexes with YY1 and RBBP7 and its downstream targets in ccRCC cells
  86. Hypereosinophilic syndrome with elevated IgG4 and T-cell clonality: A report of two cases
  87. Electroacupuncture alleviates sciatic nerve injury in sciatica rats by regulating BDNF and NGF levels, myelin sheath degradation, and autophagy
  88. Polydatin prevents cholesterol gallstone formation by regulating cholesterol metabolism via PPAR-γ signaling
  89. RNF144A and RNF144B: Important molecules for health
  90. Analysis of the detection rate and related factors of thyroid nodules in the healthy population
  91. Artesunate inhibits hepatocellular carcinoma cell migration and invasion through OGA-mediated O-GlcNAcylation of ZEB1
  92. Endovascular management of post-pancreatectomy hemorrhage caused by a hepatic artery pseudoaneurysm: Case report and review of the literature
  93. Efficacy and safety of anti-PD-1/PD-L1 antibodies in patients with relapsed refractory diffuse large B-cell lymphoma: A meta-analysis
  94. SATB2 promotes humeral fracture healing in rats by activating the PI3K/AKT pathway
  95. Overexpression of the ferroptosis-related gene, NFS1, corresponds to gastric cancer growth and tumor immune infiltration
  96. Understanding risk factors and prognosis in diabetic foot ulcers
  97. Atractylenolide I alleviates the experimental allergic response in mice by suppressing TLR4/NF-kB/NLRP3 signalling
  98. FBXO31 inhibits the stemness characteristics of CD147 (+) melanoma stem cells
  99. Immune molecule diagnostics in colorectal cancer: CCL2 and CXCL11
  100. Inhibiting CXCR6 promotes senescence of activated hepatic stellate cells with limited proinflammatory SASP to attenuate hepatic fibrosis
  101. Cadmium toxicity, health risk and its remediation using low-cost biochar adsorbents
  102. Pulmonary cryptococcosis with headache as the first presentation: A case report
  103. Solitary pulmonary metastasis with cystic airspaces in colon cancer: A rare case report
  104. RUNX1 promotes denervation-induced muscle atrophy by activating the JUNB/NF-κB pathway and driving M1 macrophage polarization
  105. Morphometric analysis and immunobiological investigation of Indigofera oblongifolia on the infected lung with Plasmodium chabaudi
  106. The NuA4/TIP60 histone-modifying complex and Hr78 modulate the Lobe2 mutant eye phenotype
  107. Experimental study on salmon demineralized bone matrix loaded with recombinant human bone morphogenetic protein-2: In vitro and in vivo study
  108. A case of IgA nephropathy treated with a combination of telitacicept and half-dose glucocorticoids
  109. Analgesic and toxicological evaluation of cannabidiol-rich Moroccan Cannabis sativa L. (Khardala variety) extract: Evidence from an in vivo and in silico study
  110. Wound healing and signaling pathways
  111. Combination of immunotherapy and whole-brain radiotherapy on prognosis of patients with multiple brain metastases: A retrospective cohort study
  112. To explore the relationship between endometrial hyperemia and polycystic ovary syndrome
  113. Research progress on the impact of curcumin on immune responses in breast cancer
  114. Biogenic Cu/Ni nanotherapeutics from Descurainia sophia (L.) Webb ex Prantl seeds for the treatment of lung cancer
  115. Dapagliflozin attenuates atrial fibrosis via the HMGB1/RAGE pathway in atrial fibrillation rats
  116. Ecology and Environmental Science
  117. Optimization and comparative study of Bacillus consortia for cellulolytic potential and cellulase enzyme activity
  118. The complete mitochondrial genome analysis of Haemaphysalis hystricis Supino, 1897 (Ixodida: Ixodidae) and its phylogenetic implications
  119. Epidemiological characteristics and risk factors analysis of multidrug-resistant tuberculosis among tuberculosis population in Huzhou City, Eastern China
  120. Indices of human impacts on landscapes: How do they reflect the proportions of natural habitats?
  121. Genetic analysis of the Siberian flying squirrel population in the northern Changbai Mountains, Northeast China: Insights into population status and conservation
  122. Diversity and environmental drivers of Suillus communities in Pinus sylvestris var. mongolica forests of Inner Mongolia
  123. Agriculture
  124. Integrated analysis of transcriptome, sRNAome, and degradome involved in the drought-response of maize Zhengdan958
  125. Variation in flower frost tolerance among seven apple cultivars and transcriptome response patterns in two contrastingly frost-tolerant selected cultivars
  126. Heritability of durable resistance to stripe rust in bread wheat (Triticum aestivum L.)
  127. Animal Science
  128. Effect of sex ratio on the life history traits of an important invasive species, Spodoptera frugiperda
  129. Plant Sciences
  130. Hairpin in a haystack: In silico identification and characterization of plant-conserved microRNA in Rafflesiaceae
  131. Widely targeted metabolomics of different tissues in Rubus corchorifolius
  132. The complete chloroplast genome of Gerbera piloselloides (L.) Cass., 1820 (Carduoideae, Asteraceae) and its phylogenetic analysis
  133. Field trial to correlate mineral solubilization activity of Pseudomonas aeruginosa and biochemical content of groundnut plants
  134. Correlation analysis between semen routine parameters and sperm DNA fragmentation index in patients with semen non-liquefaction: A retrospective study
  135. Plasticity of the anatomical traits of Rhododendron L. (Ericaceae) leaves and its implications in adaptation to the plateau environment
  136. Effects of Piriformospora indica and arbuscular mycorrhizal fungus on growth and physiology of Moringa oleifera under low-temperature stress
  137. Effects of different sources of potassium fertiliser on yield, fruit quality and nutrient absorption in “Harward” kiwifruit (Actinidia deliciosa)
  138. Comparative efficiency and residue levels of spraying programs against powdery mildew in grape varieties
  139. The DREB7 transcription factor enhances salt tolerance in soybean plants under salt stress
  140. Food Science
  141. Phytochemical analysis of Stachys iva: Discovering the optimal extract conditions and its bioactive compounds
  142. Review on role of honey in disease prevention and treatment through modulation of biological activities
  143. Computational analysis of polymorphic residues in maltose and maltotriose transporters of a wild Saccharomyces cerevisiae strain
  144. Optimization of phenolic compound extraction from Tunisian squash by-products: A sustainable approach for antioxidant and antibacterial applications
  145. Liupao tea aqueous extract alleviates dextran sulfate sodium-induced ulcerative colitis in rats by modulating the gut microbiota
  146. Toxicological qualities and detoxification trends of fruit by-products for valorization: A review
  147. Polyphenolic spectrum of cornelian cherry fruits and their health-promoting effect
  148. Optimizing the encapsulation of the refined extract of squash peels for functional food applications: A sustainable approach to reduce food waste
  149. Advancements in curcuminoid formulations: An update on bioavailability enhancement strategies curcuminoid bioavailability and formulations
  150. Impact of saline sprouting on antioxidant properties and bioactive compounds in chia seeds
  151. The dilemma of food genetics and improvement
  152. Bioengineering and Biotechnology
  153. Impact of hyaluronic acid-modified hafnium metalorganic frameworks containing rhynchophylline on Alzheimer’s disease
  154. Emerging patterns in nanoparticle-based therapeutic approaches for rheumatoid arthritis: A comprehensive bibliometric and visual analysis spanning two decades
  155. Application of CRISPR/Cas gene editing for infectious disease control in poultry
  156. Preparation of hafnium nitride-coated titanium implants by magnetron sputtering technology and evaluation of their antibacterial properties and biocompatibility
  157. Preparation and characterization of lemongrass oil nanoemulsion: Antimicrobial, antibiofilm, antioxidant, and anticancer activities
  158. Corrigendum
  159. Corrigendum to “Utilization of convolutional neural networks to analyze microscopic images for high-throughput screening of mesenchymal stem cells”
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