Home Medicine NORAD modulates miR-30c-5p-LDHA to protect lung endothelial cells damage
Article Open Access

NORAD modulates miR-30c-5p-LDHA to protect lung endothelial cells damage

  • , , , , and EMAIL logo
Published/Copyright: April 6, 2022
Download Article (PDF)
Open Medicine
From the journal Open Medicine Volume 17 Issue 1

Abstract

Acute lung injury (ALI) is a devastating human malignancy characterized by excessively uncontrolled inflammation and lung endothelial dysfunction. Non-coding RNAs play essential roles in endothelial protections during the pathological processes of ALI. The precise functions and molecular mechanisms of the lncRNA-NORAD-mediated endothelial protection remain obscure. This study reports NORAD was significantly induced in human pulmonary microvascular endothelial cells (HPMECs) under lipopolysaccharide (LPS) treatment. Silencing NORAD effectively protected HPMECs against the LPS-induced cell dysfunction. In addition, RNA pull-down and luciferase assay validated that NORAD sponged miR-30c-5p, which showed reverse functions of NORAD in the LPS-induced cell injury of HPMECs. Furthermore, the glucose metabolism of HPMECs was significantly elevated under LPS stimulation which promoted the glucose consumption and extracellular acidification rate (ECAR) of HPMECs. Inhibiting NORAD or overexpressing miR-30c-5p suppressed glucose metabolism in HPMECs, leading to protective effects on HPMECs under LPS stimulation. The glycolysis key enzyme, lactate dehydrogenase-A (LDHA), was subsequently identified as a direct target of miR-30c-5p. Finally, recovery of miR-30c-5p in NORAD-overexpressing HPMECs effectively overrode the NORAD-promoted glycolysis and impaired endothelial dysfunction under LPS stimulation by targeting LDHA. Summarily, we demonstrated a NORAD-miR-30c-5p-LDHA-glycolysis axis in the LPS-induced HPMECs dysfunction in vitro and in vivo, contributing to the development of anti-ALI therapeutic approaches.

Keywords: lncRNA NORAD; acute lung injury; HPMECs; miR-30c-5p; glucose metabolism; LDHA

1 Introduction

Acute lung injury (ALI) is a devastating human malignancy with high incidence and mortality [1]. ALI is characterized by excessive uncontrolled inflammation, lung endothelial dysfunction, and oxidative damage, leading to progress towards acute respiratory distress syndrome (ARDS) [2]. Thus, it is urgent to investigate effectively the therapeutic approaches for ALI treatment. Accumulating evidence revealed that pulmonary endothelium plays essential roles and acts as an orchestrator of ALI [3]. In the lining of pulmonary vessel, endothelial cells (ECs) are tightly connected to adjacent ones through intercellular junctions, forming a semipermeable barrier which functions in lung homoeostasis via regulating the transport of fluids, proteins, and inflammatory factors [4,5]. In response to stimuli, such as lipopolysaccharide (LPS), reactive oxygen species, or other pathological factors, barrier disruption occurs in ECs and proinflammatory proteins are secreted by injured ECs [6]. Therefore, targeting the dysregulated endothelial induced by stimuli will be a potentially effective strategy for ALI treatment.

Long non-codingRNAs (lncRNAs) are a family of RNAs with relatively larger size (longer than 200 nucleotides), and non-protein coding capacity [7]. They function as scaffolds or decoys of miRNAs or mRNAs in cytoplasm or nucleus via regulating transcriptional or posttranscriptional processes and epigenetic modifications [8]. Studies have revealed that lncRNAs are important mediators in various cellular processes including proliferation, migration, stress response, metabolism, inflammation, and cell death [9]. Moreover, research works have indicated fundamental roles of lncRNAs in acute lung inflammation and infectious diseases [10]. A recent study uncovered that lncRNA MALAT1 participated in pulmonary pathogeneses via regulating the aberrant macrophage activation [11]. MALAT1 was significantly upregulated in LPS-treated macrophages and silencing MALAT1 effectively attenuates pro-inflammatory activation of aberrant macrophages [11], suggesting targeting the lncRNA-mediated molecular pathway may contribute to treatment of ALI. LncRNA activated by DNA damage (NORAD) has been reported to be significantly upregulated in diverse human cancers and contributes to lung cancer progressions, including proliferation [12], apoptosis [12], epithelial to mesenchymal transition [13], chemoresistance [14], and migration [15]. Currently, the precise roles and molecular targets of NORAD in the LPS-stimulated dysfunction of human pulmonary microvascular endothelial cells (HPMECs) as well as ALI remain unclear.

Studies have unveiled that lncRNAs sponge target miRNAs to form ceRNA networks, leading to sequestering miRNAs [8]. Currently, the underlying molecular targets of the NORAD-regulated HPMECs dysfunction under LPS stimulation remain unclear. This study aimed to investigate the roles of NORAD in the LPS-stimulated HPMECs dysfunction. The NORAD-participated ceRNA network as well as downstream targets of miRNA will be identified. Our results will present new insights for the non-coding RNA-mediated HPMECs dysfunction, providing a potentially therapeutic approach against ALI.

2 Materials and methods

2.1 ALI patient samples collection and HPMECs isolation

Ethics of this study was approved by the Ethics Committee of Ruijin Hospital, Shanghai Jiao Tong University School of Medicine and followed the ethical principles for medical research involving human subjects outlined in the Declaration of Helsinki. Surgeries of ALI patients were carried out in Ruijin Hospital, Shanghai Jiao Tong University School of Medicine between April 1, 2018 and July 20, 2019. ALI was diagnosed based on the regular diagnostic criteria for acute lung injury. Isolation of primary HPMECs was performed according to previous reports [16]. Written informed consent was obtained from all the participants.

2.2 Cell culture

HPMECs and A549 cells were obtained from ScienCell (San Diego, CA, USA) and cultured with Dulbecco’s modified Eagle medium (DMEM; Gibco, Carlsbad, CA, USA) which contains 10% fetal bovine serum (Gibco, Carlsbad, CA, USA) and 1% penicillin/streptomycin (Invitrogen, Carlsbad, CA, USA), and incubated at 37°C with 5% CO2. LPS (Sigma-Aldrich, Shanghai, China) was dissolved in phosphate-buffered solution (PBS) and the stock solution (5 mg/mL) was stored at −20°C. HPMECs were treated with LPS at a concentration of 10 μg/mL for 24 h. Oxamate was purchased from Sigma-Aldrich (Shanghai, China). Rabbit monoclonal anti-lactate dehydrogenase-A (LDHA) (#3582), Hexokinase 2 (HK2) (#2106), and β-actin (#4970) were purchased from Cell Signaling Technology (Danvers, MA, USA).

2.3 Cell transfection of miRNAs, siRNAs, and plasmids

HPMECs (1 × 105 cells/well) were seeded in a 6-well plate and incubated for 24 h. siRNA, miRNA, plasmid DNA as well as their negative controls were transfected into cells using Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA, USA). miR-30c-5p precursor and NORAD siRNA were synthesized from GenePharma (Shanghai, China). miR-30c-5p precursor was transfected at 12.5 nM, and siRNA was transfected at 50 nM. Plasmid DNA was transfected at 2 µg/mL. 48 h post transfection, the cells were collected and subjected to downstream experiments. Experiments were performed in triplicate.

2.4 Prediction of the lncRNA-miRNA and miRNA-mRNA interactions

The binding sites of miR-30c-5p on NORAD and LDHA 3′UTR were predicted from starBase 2.0 according to previous descriptions [17].

2.5 RNA isolation and qRT-PCR

Total RNA was isolated from cells using Trizol (Sigma, Shanghai, China) according to the manufacturer’s instructions. The concentration and quality of RNA samples were measured with a spectrophotometer NanoDrop 2000 (Thermo Fisher, Waltham, MA, USA). Complementary DNA was synthesized using 1 μg RNA with a PrimeScript™ RT reagent Kit (Takara). RT-qPCR reactions were performed using a Real-time PCR Mixture assays kit (TaKaRa, Dalian, China) on a CFX96 Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). The primer sequences for qRT-PCR reactions were as follows: NORAD forward, 5′-TGATAGGATACATCTTGGACATGGA-3′ and reverse, 5′-AACCTAATGAACAAGTCCTGACATACA-3′; β-actin forward, 5′-AGCACAGAGCCTCGCCTT-3′ and reverse, 5′-CATCATCCATGGTGAGCTGG-3′; miR-30c-5p forward, 5′-AGCGTCGTATCCAGTGCAAT-3′ and reverse, 5′-GTCGTATCCAGTGCGTGTCG-3′; and U6 forward, 5′-CTCGCTTCGGCAGCACA-3′ and reverse, 5′-AACGCTTCACGAATTTGCGT-3′. Conditions were set as follows: 95°C for 30 s, 40 cycles at 95°C for 5 s, annealing, and extension at 60°C for 30 s, and melt curve analysis. U6 was used as an internal control for miR-30c-5p and human β-actin was used as an internal control for mRNA detection. The relative expression levels were calculated using the 2−ΔΔCt method. Experiments were performed in triplicate.

2.6 Luciferase reporter assays

Human wild-type (WT)- or binding site mutant (MUT)-NORAD or LDHA 3′-UTR was amplified and inserted into the pSI-Check2 vector. HPMECs and A549 cells were seeded in 96-well plates and cultured for 24 h, followed by co-transfection with control miRNA or miR-30c-5p plus WT- or MUT-NORAD or LDHA 3′UTR using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA). 48 h post transfection, firefly luciferase and Renilla luciferase activities were examined using the dual-luciferase reporter system (Luciferase Assay Reagent; Promega, San Luis Obispo, CA, USA). Experiments were performed in triplicate.

2.7 Measurements of LDH and Caspase-3 activity

The measurements of LDH and Caspase-3 activity were performed using the Lactate Dehydrogenase Activity Assay Kit (#MAK066, Sigma, Shanghai, China) and Caspase-3 Activity Assay Kit (Fluorometric) (#ab252897, Abcam, Cambridge, UK) according to the manufacturer’s instructions. Experiments were repeated three times.

2.8 RNA pull-down assay

The scramble control, sense, and antisense NORAD probes were biotin-labeled from RiboBio Co. Ltd (Guangzhou, China). Cell lysates were extracted using RIPA buffer (Sigma, Shanghai, China) and incubated with probes for 2 h. Streptavidin-coupled agarose beads (Thermo Fisher Scientific, Shanghai, China) were added into the mixture and incubated for 2 h. After washing, the miR-30c-5p was pulled down in the RNA complex. The amount of miR-30c-5p in the RNA complex was determined by qRT-PCR and Northern blot. Probes of sense and antisense NORAD are: Sense: 5′-GGUUGUAAACAGGAUGGCAUAGAGCUCUCUGCGCG-3′; Antisense: 5′-GGUGCAGAGAGCUCUAUGCCAUCCUGUUUACAGCG-3′.

2.9 Measurements of glucose metabolism

The glucose metabolism rate was evaluated by glucose consumption assay and the extracellular acidification rate (ECAR) assay using the Glucose Uptake Colorimetric Assay Kit (#MAK083) (Sigma-Aldrich, Shanghai, China) and the glycolysis stress kit (Agilent, Santa Clara, CA, USA) on a Seahorse XFp Analyzer (Agilent, Santa Clara, CA, USA) according to the manufacturer’s protocol. Results were normalized to the protein concentrations of each group. Experiments were repeated three times and performed in triplicate.

2.10 Cell viability assay

Viability of HPMECs under LPS treatment was determined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assay. Cells (1 × 105 per well) were seeded in a 24-well plate for 24 h at regular cell culture condition. After LPS treatments, 0.1 mL of MTT solution (5 mg/mL in PBS) was added to cells followed by 2 h incubation. Lysis buffer containing dimethyl sulfoxide (DMSO) was added into each well and incubated for 4 h at 37°C. The optical density value was measured at 450 nm. Experiments were performed in triplicate and repeated three times.

2.11 Cell apoptosis assay

The cell apoptosis of HPMECs under LPS treatment was determined by flowcytometry using the Annexin V-FITC Apoptosis Kit (Thermo Fisher Inc., Waltham, MA, USA) according to the manufacturer’s instructions. In brief, HPMECs (3 × 105 cells per well) were seeded into 6-well cell culture plates for 24 h. After LPS treatments, cells were collected by trypsinization and washed in suspension by cold PBS. Binding buffer (500 µL) was added into suspension and the Annexin-FITC and propidium iodide (PI) (5 µL of each) were added into each group and fully mixed. Cells were incubated for 15 min in the dark at room temperature. Analysis of cell apoptosis/death was performed using FACSCalibur flowcytometry (Becton Dickinson, San Jose, CA, USA). Experiments were repeated three times.

2.12 Western blotting

HPMECs were trypsinized and washed by PBS followed by incubation with lysis buffer and radioimmunoprecipitation buffer (Beyotime, Shanghai, China) containing protease inhibitor cocktail (Beyotime) on ice for 20 min. Protein concentration was determined using an Enhanced BCA Protein Assay Kit (Beyotime). Equal amount of protein sample (40 μg) was loaded into each well and resolved with 10% SDS-PAGE, followed by electro-transferring to a polyvinylidene fluoride (PVDF) membrane. Membranes were blocked with 5% BSA in PBST for 1 h at room temperature followed by incubation with primary antibodies (1:1,000) at 4°C overnight. The membranes were washed with TBST three times with 5 min gap between each wash. Membranes were incubated with secondary antibody for 1 h at room temperature. β-actin was a loading control. Protein expressions were detected using an ECL reagent (Santa Cruz Biotechnology). Experiments were repeated three times.

2.13 LPS-induced ALI experiments in in vivo rat model

This study was approved by the Institutional Animal Care and Use Committee of the Ruijin Hospital, Shanghai Jiao Tong University School of Medicine and in accordance with ARRIVE guidelines. All rats received regular care. Male Sprague-Dawley (SD) rats (400–450 g) were housed at 22–24°C of a 12:12 h light–dark cycle, with free access to food and water. Rats (n = 8 per group) were randomly assigned into seven groups: (1) sham, (2) LPS, (3) LPS + control shRNA, and (4) LPS + sh NORAD. Control shRNA or sh NORAD was transfected into rat lung tissues and 48 h after transfection, rats were anesthetized with 10% chloral hydrate (350 mg/kg, i.p.) and received i.p. injection of 0.9% saline solution (control) or LPS (5 mg/kg). Rats were euthanized 12 h after LPS treatment and the perfused lungs were harvested, and the lung pulmonary microvascular endothelial cells were isolated according to previous reports [18]. Cells were immediately stored in liquid nitrogen for downstream analysis.

2.14 Statistical analysis

Statistical analyses were performed using the Prism 7.0 software (GraphPad, San Diego, CA, USA). Data are expressed as the mean value ± standard deviation (mean value ± SD). Two-tailed Student’s t-test was applied for comparison of difference between two independent groups. One-way analysis of variance (ANOVA) followed by Holm-Sidak post hoc test was applied to determine significant differences among three or more groups. Experiments were conducted in triplicate. A value of p < 0.05 was considered to be statistically significant.

3 Results

3.1 NORAD is induced and involves in HPMECs death during LPS stimulation

To evaluate the roles of NORAD in the sepsis-evoked ALI, expressions of NORAD were compared in pulmonary microvascular endothelial cells from patients with ALI (n = 40) and healthy controls. Results from qRT-PCR showed that NORAD was significantly upregulated in pulmonary microvascular endothelial cells from ALI patients compared with those from controls (Figure 1a).

Figure 1 Roles of NORAD in the LPS-induced HPMEC cell injury. (a) Primary HPMECs from patient lung tissues (n = 40) and healthy controls were isolated and the expressions of NORAD were analyzed by qRT-PCR. (b) HPMECs were treated with LPS at 0, 0.5, 1, 5, 10, or 50 µg/mL. Expressions of NORAD were examined by qRT-PCR. (c) HPMECs were transfected with control siRNA or si NORAD, followed by treatment with LPS at the indicated concentrations. Cell responses were evaluated by MTT assay and (d) Caspase-3 activity assay. *, p < 0.05; **, p < 0.01; ***, p < 0.001.
Figure 1

Roles of NORAD in the LPS-induced HPMEC cell injury. (a) Primary HPMECs from patient lung tissues (n = 40) and healthy controls were isolated and the expressions of NORAD were analyzed by qRT-PCR. (b) HPMECs were treated with LPS at 0, 0.5, 1, 5, 10, or 50 µg/mL. Expressions of NORAD were examined by qRT-PCR. (c) HPMECs were transfected with control siRNA or si NORAD, followed by treatment with LPS at the indicated concentrations. Cell responses were evaluated by MTT assay and (d) Caspase-3 activity assay. *, p < 0.05; **, p < 0.01; ***, p < 0.001.

We then established an in vitro model of ALI through stimulation of HPMECs by LPS [19]. Expression of NORAD was significantly upregulated in LPS-treated HPMECs compared with control cells (Figure 1b). The qRT-PCR method for detecting NORAD and microRNA expressions were validated by Northern blot (Figure A1). To further investigate the biological functions of NORAD and the underlying mechanisms in the LPS-induced ALI/ARDS, NORAD was silenced in HPMECs (Figure A2a). NORAD knockdown without LPS stimulation did not affect cell viability. On the other way, HPMECs with low NORAD displayed attenuated cell death under LPS treatments compared with control siRNA transfected cells (Figure 1c). Expectedly, in control cells, LPS stimulation led to increased caspase-3 activity, which was antagonized by NORAD silencing in HPMECs (Figure 1d). Taken together, the above results suggest that NORAD plays important roles in the LPS-indued ALI and attenuation of NORAD could effectively protect the HPMECs injury.

3.2 LPS stimulation suppresses miR-30c-5p in HPMECs

We then investigated the potential molecular mechanisms for the NORAD-protected HPMECs injury under LPS stimulation. Accumulating studies uncovered that lncRNAs could sponge miRNAs to form a ceRNA regulatory network which downregulates miRNA expressions, resulting in de-repression of miRNA target expressions [8]. Interestingly, we detected significantly downregulated miR-30c-5p in pulmonary microvascular endothelial cells from patients with ALI (n = 40) compared with healthy controls (Figure 2a). Consistently, miR-30c-5p expressions were remarkedly suppressed in LPS-treated HPMECs (Figure 2b), suggesting miR-30c-5p might act as an ALI-preventing role. Consequently, overexpression of miR-30c-5p (Figure A2b) effectively decreased the LPS-induced HPMECs death (Figure 2c) and caspase-3 activity (Figure 2d), suggesting an invert role of miR-30c-5p in the pathological processes of ALI.

Figure 2 miR-30c-5p protects the LPS-induced apoptosis of HPMECs. (a) Primary HPMECs from patient lung tissues (n = 40) and healthy controls were isolated and the expressions of miR-30c-5p were analyzed by qRT-PCR. (b) HPMECs were treated with LPS at 0, 0.5, 1, 5, 10 or 50 µg/mL. Expressions of miR-30c-5p were examined by qRT-PCR. (c) HPMECs were transfected with control miRNA or miR-30c-5p, followed by treatment with LPS at the indicated concentrations. Cell responses were evaluated by MTT assay and (d) caspase-3 activity assay. *, p < 0.05; **, p < 0.01; ***, p < 0.001.
Figure 2

miR-30c-5p protects the LPS-induced apoptosis of HPMECs. (a) Primary HPMECs from patient lung tissues (n = 40) and healthy controls were isolated and the expressions of miR-30c-5p were analyzed by qRT-PCR. (b) HPMECs were treated with LPS at 0, 0.5, 1, 5, 10 or 50 µg/mL. Expressions of miR-30c-5p were examined by qRT-PCR. (c) HPMECs were transfected with control miRNA or miR-30c-5p, followed by treatment with LPS at the indicated concentrations. Cell responses were evaluated by MTT assay and (d) caspase-3 activity assay. *, p < 0.05; **, p < 0.01; ***, p < 0.001.

3.3 NORAD downregulates miR-30c-5p by sponging it in HPMECs

Bioinformatics analysis from the lncRNA service starBase2.0 illustrated that miRNA-30c-5p contains putative NORAD binding sites (Figure 3a). To assess whether NORAD could directly regulate miR-30c-5p expressions, HPMEC and A549 cells were transfected with control siRNA or NORAD siRNA. Expected results showed that cells with higher NORAD expressions displayed effectively attenuated miR-30c-5p expressions (Figure 3b and c). To verify the binding between lncRNA-NORAD and miR-30c-5p, RNA pull-down assay was performed. Biotin-labeled scramble, sense, or antisense NORAD probe was incubated with HPMEC and A549 cell lysate. Only biotin-labeled antisense NORAD probe could pull down enriched miR-30c-5p (Figure 3d). The endogenous miR-30c-5p was effectively pulled down by control and sense probe of NORAD (Figure 3d). Similar results were observed in A549 cells (Figure 3d). To validate whether NORAD could directly bind onto the seeding region of miR-30c-5p, HPMEC cells were co-transfected with control or miR-30c-5p or luciferase vector with insertion of WT- or binding site MUT-NORAD. As we expected, luciferase activity of HPMEC cells with WT-NORAD and miR-30c-5p co-transfection was significantly suppressed (Figure 3e and f). Meanwhile, no significant change was detected in the luciferase activity of cells which were co-transfected with miR-30c-5p or control miRNA plus MUT-NORAD (Figure 3e and f). Taken together, these results demonstrated that NORAD blocked miR-30c-5p expressions via sponging it.

Figure 3 NORAD sponges miR-30c-5p in HPMECs. (a) Prediction of the binding of miR-30c-5p on NORAD from starBase. (b) HPMECs and (c) A549 cells were transfected with control siRNA or si NORAD. Expressions of miR-30c-5p were examined by qRT-PCR. (d) RNA pull-down assay was shown in HPMECs by incubation of biotin-labeled control, sense, or antisense NORAD probe with cell lysates. Enrichment of miR-30c-5p in the RNA complex was determined by qRT-PCR and Northern blot. (e) HPMECs and (f) A549 cells were transfected with control miRNA or miR-30c-5p with luciferase vector containing WT- or MUT-NORAD. Luciferase assay was performed. **, p < 0.01; ***, p < 0.001.
Figure 3

NORAD sponges miR-30c-5p in HPMECs. (a) Prediction of the binding of miR-30c-5p on NORAD from starBase. (b) HPMECs and (c) A549 cells were transfected with control siRNA or si NORAD. Expressions of miR-30c-5p were examined by qRT-PCR. (d) RNA pull-down assay was shown in HPMECs by incubation of biotin-labeled control, sense, or antisense NORAD probe with cell lysates. Enrichment of miR-30c-5p in the RNA complex was determined by qRT-PCR and Northern blot. (e) HPMECs and (f) A549 cells were transfected with control miRNA or miR-30c-5p with luciferase vector containing WT- or MUT-NORAD. Luciferase assay was performed. **, p < 0.01; ***, p < 0.001.

3.4 Glycolysis of HPMEC was activated by LPS to induce cell apoptosis

We then investigated the cellular mechanisms for the LPS-induced cell injury of HPMECs. Previous studies reported that in an LPS-induced acute lung injury mouse model, the cellular glucose metabolism of HPMECs was significantly elevated, leading to lactate secretion [20], suggesting targeting the glycolysis might contribute to protecting the LPS-induced HPMECs injury. We then evaluated the glycolysis rate of HPMECs under LPS treatments. Results in Figure 4a and b showed that HPMECs with LPS treatment displayed significantly increased glucose uptake and ECAR. Moreover, the glycolysis key enzymes, HK2, and LDHA were significantly upregulated under LPS treatments (Figure 4c). To verify whether LPS induced HPMECs injury through activating the glycolysis, the HPMECs were pre-treated with glycolysis inhibitor, Oxamate for 48 h, followed by LPS stimulation. Expectedly, blocking glycolysis effectively prevented the cell apoptosis induced by LPS (Figure 4d). Consistently, the LDH release and activity of caspase-3 were inhibited with glycolysis inhibitor under LPS treatments (Figure 4e and f). In summary, our results suggest that blocking the LPS-induced glycolysis might be an effective approach to protect ALI.

Figure 4 Glycolysis rate of HPMECs is elevated by LPS stimulation. (a) HPMECs were treated with control or LPS at 5 µg/mL. The glucose consumption, (b) ECAR and (c) glycolysis enzymes were examined. (d) HPMECs were treated with control or Oxamate at 5 mM, followed by treatments with LPS at the indicated concentrations. Cell viability was examined by MTT assay. (e and f) HPMECs were treated with LPS alone or with Oxamate at the indicated concentrations. The LDH activity and Caspase-3 activity were examined. *, p < 0.05; **, p < 0.01.
Figure 4

Glycolysis rate of HPMECs is elevated by LPS stimulation. (a) HPMECs were treated with control or LPS at 5 µg/mL. The glucose consumption, (b) ECAR and (c) glycolysis enzymes were examined. (d) HPMECs were treated with control or Oxamate at 5 mM, followed by treatments with LPS at the indicated concentrations. Cell viability was examined by MTT assay. (e and f) HPMECs were treated with LPS alone or with Oxamate at the indicated concentrations. The LDH activity and Caspase-3 activity were examined. *, p < 0.05; **, p < 0.01.

3.5 Reverse effects of NORAD and miR-30c-5p in modulating anaerobic glycolysis

The above results revealed reverse roles of NORAD and miR-30c-5p in the LPS-induced HPMECs injury. To investigate whether NORAD and miR-30c-5p modulate glycolysis in an invert pattern, effects of NORAD silencing and miR-30c-5p overexpression on glucose metabolism were assessed. Results in Figure 5a demonstrated that silencing NORAD significantly suppressed glucose uptake. In addition, the lactate product was inhibited in NORAD siRNA transfected cells (Figure 5b). Consistently, overexpression of miR-30c-5p remarkedly blocked the glycolysis rate of HPMECs (Figure 5c and d).

Figure 5 Invert roles of NORAD and miR-30c-5p in glycolysis. (a) The glucose consumption and (b) ECAR were examined in HPMECs without or with NORAD silencing. (c) The glucose consumption and (d) ECAR were examined in HPMECs without or with miR-30c-5p overexpression. *, p < 0.05; **, p < 0.01.
Figure 5

Invert roles of NORAD and miR-30c-5p in glycolysis. (a) The glucose consumption and (b) ECAR were examined in HPMECs without or with NORAD silencing. (c) The glucose consumption and (d) ECAR were examined in HPMECs without or with miR-30c-5p overexpression. *, p < 0.05; **, p < 0.01.

3.6 miR-30c-5p targets LDHA to attenuate glycolysis of HPMECs

Accumulating studies described miRNAs function through binding to the 3′UTR of their target mRNAs to interfere the mRNA stability [21]. We thus investigated the targets of miR-30c-5p in the LPS-induced lung endothelial injury. Targets of miR-30c-5p were predicted from starBase2.0 non-coding RNA service. We observed LDHA (Figure 6a), which is a glycolysis key enzyme that converts the pyruvate to lactate, is a potential target of miR-30c-5p. Consistent results showed that LDHA was significantly upregulated in pulmonary microvascular endothelial cells from patients with ALI (n = 40) compared with healthy controls (Figure A3), suggesting LDHA is positively associated with ALI. Contrarily, we detected that overexpression of miR-30c-5p significantly downregulated LDHA protein expression in HPMECs (Figure 6b). To obtain the direct evidence of miR-30c-5p–LDHA interaction, luciferase reporter assays were performed. HPMEC cells were co-transfected with control or miR-30c-5p or luciferase vector with insertion of WT or binding site MUT 3′UTR of LDHA. Expectedly, luciferase activity of HPMEC cells with WT-LDHA 3′UTR and miR-30c-5p co-transfection was significantly suppressed but no significant change was detected in the luciferase activity of cells which were co-transfected with miR-30c-5p or control miRNA plus Mut-LDHA 3′UTR (Figure 6c).

Figure 6 miR-30c-5p directly targets LDHA in HPMECs. (a) Prediction of the binding of miR-30c-5p on 3′UTR of LDHA from starBase. (b) HPMECs were transfected with control miRNA or miR-30c-5p. Expressions of LDHA were examined by western blot. (c) HPMECs were transfected with control miRNA or miR-30c-5p with luciferase vector containing WT- or MUT-LDHA 3′UTR. Luciferase assay was performed. (d) HPMECs were transfected with control miRNA, miR-30c-5p alone, or with LDHA. Expressions of LDHA were examined by western blot. (e) The glucose consumption and (f) ECAR were examined in the above transfected cells. (g) The transfected cells were treated with LPS at the indicated concentrations. Cell survival rates were determined by MTT assay and (h) Annexin V apoptosis assay. *, p < 0.05; **, p < 0.01.
Figure 6

miR-30c-5p directly targets LDHA in HPMECs. (a) Prediction of the binding of miR-30c-5p on 3′UTR of LDHA from starBase. (b) HPMECs were transfected with control miRNA or miR-30c-5p. Expressions of LDHA were examined by western blot. (c) HPMECs were transfected with control miRNA or miR-30c-5p with luciferase vector containing WT- or MUT-LDHA 3′UTR. Luciferase assay was performed. (d) HPMECs were transfected with control miRNA, miR-30c-5p alone, or with LDHA. Expressions of LDHA were examined by western blot. (e) The glucose consumption and (f) ECAR were examined in the above transfected cells. (g) The transfected cells were treated with LPS at the indicated concentrations. Cell survival rates were determined by MTT assay and (h) Annexin V apoptosis assay. *, p < 0.05; **, p < 0.01.

We then asked whether the miR-30c-5p-mediated HPMECs protection was through targeting LDHA. Rescue experiments were performed by transfection of control miRNA, miR-30c-5p alone, or plus LDHA overexpression vector into HPMEC cells. Results from western blot showed the combined transfection of miR-30c-5p with LDHA successfully recovered the LDHA protein expression (Figure 6d). Furthermore, rescue of LDHA in miR-30c-5p-overexpressing HPMEC cells effectively restored the glucose uptake (Figure 6e) and ECAR (Figure 6f). Importantly, HPMEC cells with recovery of LDHA exhibited obviously increased sensitization to LPS by cell viability assay (Figure 6g) and apoptosis assay (Figure 6h). Taken together, the above data consistently demonstrated that miR-30c-5p protects HPMECs under LPS-induced cell injury via direct targeting of LDHA-glycolysis pathway.

3.7 Blocking NORAD protects LPS-induced cell death through the miR-30c-5p-LDHA axis

Finally, we investigated whether the NORAD-modulated HPMECs protection under LPS treatment was through the miR-30c-5p-LDHA axis. Mechanism rescue experiments were conducted by co-transfection of control vector, NORAD overexpression vector alone, or plus miR-30c-5p into HPMECs. Western blot results showed overexpression of NORAD significantly suppressed miR-30c-5p (Figure 7a) and upregulated LDHA protein expressions (Figure 7b). Recovery of miR-30c-5p effectively re-suppressed LDHA expression (Figure 7a and b). Consistently, restoration of miR-30c-5p in NORAD-overexpressing HPMECs successfully recovered the glucose consumption (Figure 7c) and ECAR (Figure 7d) compared with NORAD transfection alone. Moreover, restoration of miR-30c-5p in NORAD-overexpressing HPMECs effectively protected cells under the LPS-induced apoptosis by cell viability assay (Figure 7e) and Annexin V apoptosis assay (Figure 7f).

Figure 7 Roles of the NORAD-miR-30c-5p-LDHA axis in the LPS-induced HPMECs dysfunction. (a) HPMECs were transfected with control, NORAD alone, or with miR-30c-5p. Expressions of miR-30c-5p and (b) LDHA were examined by qRT-PCR and western blot, respectively. (c) The glucose consumption and (d) ECAR were examined in the above transfected cells. (e) HPMECs were transfected with control, NORAD, NORAD plus miR-30c-5p were treated with control or LPS. Cell survival rates were determined by MTT assay and (f) Annexin V apoptosis assay. *, p < 0.05; **, p < 0.01.
Figure 7

Roles of the NORAD-miR-30c-5p-LDHA axis in the LPS-induced HPMECs dysfunction. (a) HPMECs were transfected with control, NORAD alone, or with miR-30c-5p. Expressions of miR-30c-5p and (b) LDHA were examined by qRT-PCR and western blot, respectively. (c) The glucose consumption and (d) ECAR were examined in the above transfected cells. (e) HPMECs were transfected with control, NORAD, NORAD plus miR-30c-5p were treated with control or LPS. Cell survival rates were determined by MTT assay and (f) Annexin V apoptosis assay. *, p < 0.05; **, p < 0.01.

To further validate the above in vitro conclusions, we performed in vivo experiments by establishing an ALI rat model. Control or LPS was injected i.p. to induce ALI in rats 48 h after transfection of control shRNA or sh NORAD. Rat lung endothelial cells were isolated. Consistent with in vitro results, rat lung endothelial cells in LPS treatment group exhibited increased NORAD and LDHA expression and decreased miR-30c-5p expression (Figure 8a–c). Furthermore, silencing NORAD effectively recovered miR-30c-5p expression and downregulated LDHA expression in rat lung microvascular endothelial cells under LPS treatments (Figure 8a–c). Generally, these in vitro and in vivo results consistently validated that the NORAD-promoted HPMECs cell death under LPS through modulating the miR-30c-5p-LDHA axis (Figure 9).

Figure 8 Roles of NORAD-miR-30c-5p-LDHA axis in an ALI rat model. (a) Control or LPS was injected i.p. to induced ALI in rats 48 h after transfection of control shRNA or sh NORAD. Rat lung endothelial cells were isolated. Expressions of NORAD, (b) miR-30c-5p and (c) LDHA mRNA were examined. *, p < 0.05; **, p < 0.01; ***, p < 0.001.
Figure 8

Roles of NORAD-miR-30c-5p-LDHA axis in an ALI rat model. (a) Control or LPS was injected i.p. to induced ALI in rats 48 h after transfection of control shRNA or sh NORAD. Rat lung endothelial cells were isolated. Expressions of NORAD, (b) miR-30c-5p and (c) LDHA mRNA were examined. *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Figure 9 A working model for the NORAD-miR-30c-5p-LDHA axis in the LPS-induced HPMECs dysfunction.
Figure 9

A working model for the NORAD-miR-30c-5p-LDHA axis in the LPS-induced HPMECs dysfunction.

4 Discussion

ALI is a devastating human malignancy which is characterized by excessively uncontrolled inflammation, lung epithelial and endothelial cell dysfunction, and oxidative damage [1]. Thus, it is urgent to investigate effectively therapeutic approaches for ALI treatment. Accumulating evidence revealed that pulmonary endothelium plays essential roles which contribute to the development and progression of ALI [2]. The LPS-stimulated HPMECs has been widely used as an in vitro ALI model to study the effects and molecular mechanisms of lung endothelial cell dysfunction [6]. NORAD has been reported to be highly expressed in lung tissues and is associated with tumorigenesis and progresses of lung cancer [14,22]. In this study, we reported a lncRNA NORAD-regulated endothelial cell dysfunction under LPS stimulation using an in vitro HPMECs model and in vivo rat LPS model. NORAD was observed to be significantly induced under LPS treatment. Expectedly, silencing NORAD effectively protected HPMECs against the LPS-induced HPMECs apoptosis. These results highlighted that NORAD is a potentially therapeutic target in management of ALI.

Recent studies uncovered that lncRNAs function as sponges of miRNAs to suppress their expressions, leading to re-activation of downstream target mRNAs [8]. To explore the molecular targets of NORAD, we identified miR-30c-5p as a direct target of NORAD which significantly downregulated miR-30c-5p expression in HPMECs. RNA pull-down assay and luciferase assay consistently demonstrated that NORAD was associated with miR-30c-5p to form a ceRNA regulatory network in lung endothelial cells. miR-30c-5p is a member of miR-30 family which is conservely expressed in lung tissues [23]. In this study, we detected miR-30c-5p was suppressed during the LPS-stimulated lung endothelial cell death, suggesting miR-30c-5p plays a protective role in ALI.

Accumulating studies revealed that a series of pathophysiological alterations such as endothelial cell glucose metabolism are associated with ALI-induced lung sepsis [14]. During ALI, hyperlactatemia occurs due to the inadequate oxygen supply-activated upregulation of anaerobic glycolysis [24]. Anaerobic glycolysis is the metabolization of glucose to lactate controlled by multiple speed-limiting glycolytic enzymes such as LDHA [25]. A recent study showed blocking glycolysis pathway by inhibitor, PFKFB3, 3-(3-pyridinyl)-1-(4-pyridinyl)-2-propen-1-one(3PO), which led to decreased glucose uptake and suppression of glycolytic flux to lactate, was an effective approach to prevent the sepsis-related ALI [24], indicating inhibition of cellular glycolysis is a potential way to treat ALI. In this study, LPS exposure remarkedly stimulated the glycolysis rate of HPMECs, the glucose consumption, and ECAR as well as the glycolysis enzymes were significantly promoted. We then asked whether the NORAD-miR-30c-5p axis-modulated lung endothelial cells dysfunction was through targeting the LDHA-glycolysis pathway. Rescue experiments showed that recovery of miR-30c-5p in NORAD-overexpressing HPMECs cells effectively overrode the NORAD-promoted glucose metabolism and protected the LPS-induced lung endothelial cell death.

In conclusion, the current study demonstrated that inhibition of NORAD effectively ameliorated lung endothelial cell damage by LPS stimulation through modulating the miR-30c-5p-LDHA-anearobic glycolysis pathway.

tel: +86-13501747906

Acknowledgements

None.

  1. Funding information: This work was supported by grants from (a) the Shanghai Three-Year Plan of the Key Subjects Construction in Public Health-Infectious Diseases and Pathogenic Microorganism (15GWZK0102); (b) the Natural Science Foundation of Shanghai (17ZR1415900), and (c) Medical Guidance Project of Shanghai Science and Technology Commission (No. 18411966400 and No. 16411970700).

  2. Author contributions: Y.H.Z. and E.Q.M. designed the experiments. Y.H.Z., C.Y.C., Q.T.L., H.Q.S., and X.K.G. carried out the experiments and analyzed the data. Y.H.Z. and E.Q.M. wrote the manuscript. All authors participated in the manuscript preparation.

  3. Conflict of interest: The authors declared no potential conflicts of interest.

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

Appendix

Figure A1 Northern blot results of NORAD and miR-30c-5p expressions. Isolated primary pulmonary microvascular endothelial cells and HPMECs cell line were subjected to Northern blot analysis. U6 and GAPDH were internal controls for miR-30c-5p and NORAD.
Figure A1

Northern blot results of NORAD and miR-30c-5p expressions. Isolated primary pulmonary microvascular endothelial cells and HPMECs cell line were subjected to Northern blot analysis. U6 and GAPDH were internal controls for miR-30c-5p and NORAD.

Figure A2 Transfection efficiency of si NORAD and miR-30c-5p in HPMECs. (a) HPMECs were transfected with control siRNA or si NORAD. Expressions of NORAD were examined by qRT-PCR. (b) HPMECs were transfected with control miRNA or miR-30c-5p. Expressions of miR-30c-5p were examined by qRT-PCR. ***, p < 0.001.
Figure A2

Transfection efficiency of si NORAD and miR-30c-5p in HPMECs. (a) HPMECs were transfected with control siRNA or si NORAD. Expressions of NORAD were examined by qRT-PCR. (b) HPMECs were transfected with control miRNA or miR-30c-5p. Expressions of miR-30c-5p were examined by qRT-PCR. ***, p < 0.001.

Figure A3 Expression of LHDA in ALI. Primary human pulmonary microvascular endothelial cells from patient lung tissues (n = 40) and healthy controls were isolated and the expressions of LDHA were analyzed by qRT-PCR.
Figure A3

Expression of LHDA in ALI. Primary human pulmonary microvascular endothelial cells from patient lung tissues (n = 40) and healthy controls were isolated and the expressions of LDHA were analyzed by qRT-PCR.

References

[1] Mowery NT, Terzian WTH, Nelson AC. Acute lung injury. Curr Probl Surg. 2020;57:100777.10.1016/j.cpsurg.2020.100777Search in Google Scholar PubMed

[2] Butt Y, Kurdowska A, Allen TC. Acute lung injury: a clinical and molecular review. Arch Pathol Lab Med. 2016;140:345–50.10.5858/arpa.2015-0519-RASearch in Google Scholar PubMed

[3] Niethamer TK, Stabler CT, Leach JP, Zepp JA, Morley MP, Babu A, et al. Defining the role of pulmonary endothelial cell heterogeneity in response to acute lung injury. Elife. 2020;9:e53072.10.7554/eLife.53072Search in Google Scholar PubMed PubMed Central

[4] Marichal T. Endothelial cells instruct macrophages on how to respond to lung injury. Nat Immunol. 2020;21:1317–8.10.1038/s41590-020-00806-zSearch in Google Scholar PubMed

[5] Jones JH, Minshall RD. Lung endothelial transcytosis. Compr Physiol. 2020;10:491–508.10.1002/cphy.c190012Search in Google Scholar PubMed

[6] Nova Z, Skovierova H, Calkovska A. Alveolar-capillary membrane-related pulmonary cells as a target in endotoxin-induced acute lung injury. Int J Mol Sci. 2019;20:831.10.3390/ijms20040831Search in Google Scholar PubMed PubMed Central

[7] Kopp F, Mendell JT. Functional classification and experimental dissection of long noncoding RNAs. Cell. 2018;172:393–407.10.1016/j.cell.2018.01.011Search in Google Scholar PubMed PubMed Central

[8] Zhao Z, Sun W, Guo Z, Zhang J, Yu H, Liu B. Mechanisms of lncRNA/microRNA interactions in angiogenesis. Life Sci. 2020;254:116900.10.1016/j.lfs.2019.116900Search in Google Scholar PubMed

[9] Ali T, Grote P. Beyond the RNA-dependent function of LncRNA genes. Elife. 2020;9:e60583.10.7554/eLife.60583Search in Google Scholar PubMed PubMed Central

[10] Chen C, He Y, Feng Y, Hong W, Luo G, Ye Z. Long non-coding RNA review and implications in acute lung inflammation. Life Sci. 2021;269:119044.10.1016/j.lfs.2021.119044Search in Google Scholar PubMed PubMed Central

[11] Liu Y, Wang X, Li P, Zhao Y, Yang L, Yu W, et al. Targeting MALAT1 and miRNA-181a-5p for the intervention of acute lung injury/acute respiratory distress syndrome. Respir Med. 2020;175:106210.10.1016/j.rmed.2020.106210Search in Google Scholar PubMed PubMed Central

[12] Li J, Xu X, Wei C, Liu L, Wang T. Long noncoding RNA NORAD regulates lung cancer cell proliferation, apoptosis, migration, and invasion by the miR-30a-5p/ADAM19 axis. Int J Clin Exp Pathol. 2020;13:1–13.Search in Google Scholar

[13] Chen Z, Che Q, Xie C. NORAD regulates epithelial-mesenchymal transition of non-small cell lung cancer cells via miR-422a. Mol Med Rep. 2021;23:111.10.3892/mmr.2020.11750Search in Google Scholar PubMed PubMed Central

[14] Huang Q, Xing S, Peng A, Yu Z. NORAD accelerates chemo-resistance of non-small-cell lung cancer via targeting at miR-129-1-3p/SOX4 axis. Biosci Rep. 2020;40:BSR20193489.10.1042/BSR20193489Search in Google Scholar PubMed PubMed Central

[15] Wang C, Wu D, He M, Guan L, Bai D, Liang B. LncRNA NORAD accelerates the progression of non-small cell lung cancer via targeting miRNA-455/CDK14 axis. Minerva Med. 2021. 10.23736/S0026-4806.21.07194-9, Online ahead of print.Search in Google Scholar PubMed

[16] Mackay LS, Dodd S, Dougall IG, Tomlinson W, Lordan J, Fisher AJ, et al. Isolation and characterisation of human pulmonary microvascular endothelial cells from patients with severe emphysema. Respir Res. 2013;14:23.10.1186/1465-9921-14-23Search in Google Scholar PubMed PubMed Central

[17] Li JH, Liu S, Zhou H, Qu LH, Yang JH. starBase v2.0: decoding miRNA-ceRNA, miRNA-ncRNA and protein-RNA interaction networks from large-scale CLIP-Seq data. Nucleic Acids Res. 2014;42(Database issue):D92–7.10.1093/nar/gkt1248Search in Google Scholar PubMed PubMed Central

[18] Wertheim BM, Lin YD, Zhang YY, Samokhin AO, Alba GA, Arons E, et al. Isolating pulmonary microvascular endothelial cells ex vivo: Implications for pulmonary arterial hypertension, and a caution on the use of commercial biomaterials. PLoS One. 2019;14:e0211909.10.1371/journal.pone.0211909Search in Google Scholar PubMed PubMed Central

[19] Menden H, Tate E, Hogg N, Sampath V. LPS-mediated endothelial activation in pulmonary endothelial cells: role of Nox2-dependent IKK-beta phosphorylation. Am J Physiol Lung Cell Mol Physiol. 2013;304:L445–55.10.1152/ajplung.00261.2012Search in Google Scholar PubMed PubMed Central

[20] Zhong WJ, Yang HH, Guan XX, Xiong JB, Sun CC, Zhang CY, et al. Inhibition of glycolysis alleviates lipopolysaccharide-induced acute lung injury in a mouse model. J Cell Physiol. 2019;234:4641–54.10.1002/jcp.27261Search in Google Scholar PubMed

[21] Lu TX, Rothenberg ME. MicroRNA. J Allergy Clin Immunol. 2018;141:1202–7.10.1016/j.jaci.2017.08.034Search in Google Scholar PubMed PubMed Central

[22] Chen T, Qin S, Gu Y, Pan H, Bian D. Long non-coding RNA NORAD promotes the occurrence and development of non-small cell lung cancer by adsorbing MiR-656-3p. Mol Genet Genomic Med. 2019;7:e757.10.1002/mgg3.757Search in Google Scholar PubMed PubMed Central

[23] Song K, Jiang Y, Zhao Y, Xie Y, Zhou J, Yu W, et al. Members of the miR-30 family inhibit the epithelial-to-mesenchymal transition of non-small-cell lung cancer cells by suppressing XB130 expression levels. Oncol Lett. 2020;20:68.10.3892/ol.2020.11929Search in Google Scholar PubMed PubMed Central

[24] Wang L, Cao Y, Gorshkov B, Zhou Y, Yang Q, Xu J, et al. Ablation of endothelial PFKFB3 protects mice from acute lung injury in LPS-induced endotoxemia. Pharmacol Res. 2019;146:104292.10.1016/j.phrs.2019.104292Search in Google Scholar PubMed PubMed Central

[25] Koukourakis MI, Giatromanolaki A. Warburg effect, lactate dehydrogenase, and radio/chemotherapy efficacy. Int J Radiat Biol. 2019;95:408–26.10.1080/09553002.2018.1490041Search in Google Scholar PubMed

Received: 2021-09-06
Revised: 2022-01-12
Accepted: 2022-01-27
Published Online: 2022-04-06

© 2022 Yuhua Zhou et al., published by De Gruyter

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

Articles in the same Issue

  1. Research Articles
  2. AMBRA1 attenuates the proliferation of uveal melanoma cells
  3. A ceRNA network mediated by LINC00475 in papillary thyroid carcinoma
  4. Differences in complications between hepatitis B-related cirrhosis and alcohol-related cirrhosis
  5. Effect of gestational diabetes mellitus on lipid profile: A systematic review and meta-analysis
  6. Long noncoding RNA NR2F1-AS1 stimulates the tumorigenic behavior of non-small cell lung cancer cells by sponging miR-363-3p to increase SOX4
  7. Promising novel biomarkers and candidate small-molecule drugs for lung adenocarcinoma: Evidence from bioinformatics analysis of high-throughput data
  8. Plasmapheresis: Is it a potential alternative treatment for chronic urticaria?
  9. The biomarkers of key miRNAs and gene targets associated with extranodal NK/T-cell lymphoma
  10. Gene signature to predict prognostic survival of hepatocellular carcinoma
  11. Effects of miRNA-199a-5p on cell proliferation and apoptosis of uterine leiomyoma by targeting MED12
  12. Does diabetes affect paraneoplastic thrombocytosis in colorectal cancer?
  13. Is there any effect on imprinted genes H19, PEG3, and SNRPN during AOA?
  14. Leptin and PCSK9 concentrations are associated with vascular endothelial cytokines in patients with stable coronary heart disease
  15. Pericentric inversion of chromosome 6 and male fertility problems
  16. Staple line reinforcement with nebulized cyanoacrylate glue in laparoscopic sleeve gastrectomy: A propensity score-matched study
  17. Retrospective analysis of crescent score in clinical prognosis of IgA nephropathy
  18. Expression of DNM3 is associated with good outcome in colorectal cancer
  19. Activation of SphK2 contributes to adipocyte-induced EOC cell proliferation
  20. CRRT influences PICCO measurements in febrile critically ill patients
  21. SLCO4A1-AS1 mediates pancreatic cancer development via miR-4673/KIF21B axis
  22. lncRNA ACTA2-AS1 inhibits malignant phenotypes of gastric cancer cells
  23. circ_AKT3 knockdown suppresses cisplatin resistance in gastric cancer
  24. Prognostic value of nicotinamide N-methyltransferase in human cancers: Evidence from a meta-analysis and database validation
  25. GPC2 deficiency inhibits cell growth and metastasis in colon adenocarcinoma
  26. A pan-cancer analysis of the oncogenic role of Holliday junction recognition protein in human tumors
  27. Radiation increases COL1A1, COL3A1, and COL1A2 expression in breast cancer
  28. Association between preventable risk factors and metabolic syndrome
  29. miR-29c-5p knockdown reduces inflammation and blood–brain barrier disruption by upregulating LRP6
  30. Cardiac contractility modulation ameliorates myocardial metabolic remodeling in a rabbit model of chronic heart failure through activation of AMPK and PPAR-α pathway
  31. Quercitrin protects human bronchial epithelial cells from oxidative damage
  32. Smurf2 suppresses the metastasis of hepatocellular carcinoma via ubiquitin degradation of Smad2
  33. circRNA_0001679/miR-338-3p/DUSP16 axis aggravates acute lung injury
  34. Sonoclot’s usefulness in prediction of cardiopulmonary arrest prognosis: A proof of concept study
  35. Four drug metabolism-related subgroups of pancreatic adenocarcinoma in prognosis, immune infiltration, and gene mutation
  36. Decreased expression of miR-195 mediated by hypermethylation promotes osteosarcoma
  37. LMO3 promotes proliferation and metastasis of papillary thyroid carcinoma cells by regulating LIMK1-mediated cofilin and the β-catenin pathway
  38. Cx43 upregulation in HUVECs under stretch via TGF-β1 and cytoskeletal network
  39. Evaluation of menstrual irregularities after COVID-19 vaccination: Results of the MECOVAC survey
  40. Histopathologic findings on removed stomach after sleeve gastrectomy. Do they influence the outcome?
  41. Analysis of the expression and prognostic value of MT1-MMP, β1-integrin and YAP1 in glioma
  42. Optimal diagnosis of the skin cancer using a hybrid deep neural network and grasshopper optimization algorithm
  43. miR-223-3p alleviates TGF-β-induced epithelial-mesenchymal transition and extracellular matrix deposition by targeting SP3 in endometrial epithelial cells
  44. Clinical value of SIRT1 as a prognostic biomarker in esophageal squamous cell carcinoma, a systematic meta-analysis
  45. circ_0020123 promotes cell proliferation and migration in lung adenocarcinoma via PDZD8
  46. miR-22-5p regulates the self-renewal of spermatogonial stem cells by targeting EZH2
  47. hsa-miR-340-5p inhibits epithelial–mesenchymal transition in endometriosis by targeting MAP3K2 and inactivating MAPK/ERK signaling
  48. circ_0085296 inhibits the biological functions of trophoblast cells to promote the progression of preeclampsia via the miR-942-5p/THBS2 network
  49. TCD hemodynamics findings in the subacute phase of anterior circulation stroke patients treated with mechanical thrombectomy
  50. Development of a risk-stratification scoring system for predicting risk of breast cancer based on non-alcoholic fatty liver disease, non-alcoholic fatty pancreas disease, and uric acid
  51. Tollip promotes hepatocellular carcinoma progression via PI3K/AKT pathway
  52. circ_0062491 alleviates periodontitis via the miR-142-5p/IGF1 axis
  53. Human amniotic fluid as a source of stem cells
  54. lncRNA NONRATT013819.2 promotes transforming growth factor-β1-induced myofibroblastic transition of hepatic stellate cells by miR24-3p/lox
  55. NORAD modulates miR-30c-5p-LDHA to protect lung endothelial cells damage
  56. Idiopathic pulmonary fibrosis telemedicine management during COVID-19 outbreak
  57. Risk factors for adverse drug reactions associated with clopidogrel therapy
  58. Serum zinc associated with immunity and inflammatory markers in Covid-19
  59. The relationship between night shift work and breast cancer incidence: A systematic review and meta-analysis of observational studies
  60. LncRNA expression in idiopathic achalasia: New insight and preliminary exploration into pathogenesis
  61. Notoginsenoside R1 alleviates spinal cord injury through the miR-301a/KLF7 axis to activate Wnt/β-catenin pathway
  62. Moscatilin suppresses the inflammation from macrophages and T cells
  63. Zoledronate promotes ECM degradation and apoptosis via Wnt/β-catenin
  64. Epithelial-mesenchymal transition-related genes in coronary artery disease
  65. The effect evaluation of traditional vaginal surgery and transvaginal mesh surgery for severe pelvic organ prolapse: 5 years follow-up
  66. Repeated partial splenic artery embolization for hypersplenism improves platelet count
  67. Low expression of miR-27b in serum exosomes of non-small cell lung cancer facilitates its progression by affecting EGFR
  68. Exosomal hsa_circ_0000519 modulates the NSCLC cell growth and metastasis via miR-1258/RHOV axis
  69. miR-455-5p enhances 5-fluorouracil sensitivity in colorectal cancer cells by targeting PIK3R1 and DEPDC1
  70. The effect of tranexamic acid on the reduction of intraoperative and postoperative blood loss and thromboembolic risk in patients with hip fracture
  71. Isocitrate dehydrogenase 1 mutation in cholangiocarcinoma impairs tumor progression by sensitizing cells to ferroptosis
  72. Artemisinin protects against cerebral ischemia and reperfusion injury via inhibiting the NF-κB pathway
  73. A 16-gene signature associated with homologous recombination deficiency for prognosis prediction in patients with triple-negative breast cancer
  74. Lidocaine ameliorates chronic constriction injury-induced neuropathic pain through regulating M1/M2 microglia polarization
  75. MicroRNA 322-5p reduced neuronal inflammation via the TLR4/TRAF6/NF-κB axis in a rat epilepsy model
  76. miR-1273h-5p suppresses CXCL12 expression and inhibits gastric cancer cell invasion and metastasis
  77. Clinical characteristics of pneumonia patients of long course of illness infected with SARS-CoV-2
  78. circRNF20 aggravates the malignancy of retinoblastoma depending on the regulation of miR-132-3p/PAX6 axis
  79. Linezolid for resistant Gram-positive bacterial infections in children under 12 years: A meta-analysis
  80. Rack1 regulates pro-inflammatory cytokines by NF-κB in diabetic nephropathy
  81. Comprehensive analysis of molecular mechanism and a novel prognostic signature based on small nuclear RNA biomarkers in gastric cancer patients
  82. Smog and risk of maternal and fetal birth outcomes: A retrospective study in Baoding, China
  83. Let-7i-3p inhibits the cell cycle, proliferation, invasion, and migration of colorectal cancer cells via downregulating CCND1
  84. β2-Adrenergic receptor expression in subchondral bone of patients with varus knee osteoarthritis
  85. Possible impact of COVID-19 pandemic and lockdown on suicide behavior among patients in Southeast Serbia
  86. In vitro antimicrobial activity of ozonated oil in liposome eyedrop against multidrug-resistant bacteria
  87. Potential biomarkers for inflammatory response in acute lung injury
  88. A low serum uric acid concentration predicts a poor prognosis in adult patients with candidemia
  89. Antitumor activity of recombinant oncolytic vaccinia virus with human IL2
  90. ALKBH5 inhibits TNF-α-induced apoptosis of HUVECs through Bcl-2 pathway
  91. Risk prediction of cardiovascular disease using machine learning classifiers
  92. Value of ultrasonography parameters in diagnosing polycystic ovary syndrome
  93. Bioinformatics analysis reveals three key genes and four survival genes associated with youth-onset NSCLC
  94. Identification of autophagy-related biomarkers in patients with pulmonary arterial hypertension based on bioinformatics analysis
  95. Protective effects of glaucocalyxin A on the airway of asthmatic mice
  96. Overexpression of miR-100-5p inhibits papillary thyroid cancer progression via targeting FZD8
  97. Bioinformatics-based analysis of SUMOylation-related genes in hepatocellular carcinoma reveals a role of upregulated SAE1 in promoting cell proliferation
  98. Effectiveness and clinical benefits of new anti-diabetic drugs: A real life experience
  99. Identification of osteoporosis based on gene biomarkers using support vector machine
  100. Tanshinone IIA reverses oxaliplatin resistance in colorectal cancer through microRNA-30b-5p/AVEN axis
  101. miR-212-5p inhibits nasopharyngeal carcinoma progression by targeting METTL3
  102. Association of ST-T changes with all-cause mortality among patients with peripheral T-cell lymphomas
  103. LINC00665/miRNAs axis-mediated collagen type XI alpha 1 correlates with immune infiltration and malignant phenotypes in lung adenocarcinoma
  104. The perinatal factors that influence the excretion of fecal calprotectin in premature-born children
  105. Effect of femoral head necrosis cystic area on femoral head collapse and stress distribution in femoral head: A clinical and finite element study
  106. Does the use of 3D-printed cones give a chance to postpone the use of megaprostheses in patients with large bone defects in the knee joint?
  107. lncRNA HAGLR modulates myocardial ischemia–reperfusion injury in mice through regulating miR-133a-3p/MAPK1 axis
  108. Protective effect of ghrelin on intestinal I/R injury in rats
  109. In vivo knee kinematics of an innovative prosthesis design
  110. Relationship between the height of fibular head and the incidence and severity of knee osteoarthritis
  111. lncRNA WT1-AS attenuates hypoxia/ischemia-induced neuronal injury during cerebral ischemic stroke via miR-186-5p/XIAP axis
  112. Correlation of cardiac troponin T and APACHE III score with all-cause in-hospital mortality in critically ill patients with acute pulmonary embolism
  113. LncRNA LINC01857 reduces metastasis and angiogenesis in breast cancer cells via regulating miR-2052/CENPQ axis
  114. Endothelial cell-specific molecule 1 (ESM1) promoted by transcription factor SPI1 acts as an oncogene to modulate the malignant phenotype of endometrial cancer
  115. SELENBP1 inhibits progression of colorectal cancer by suppressing epithelial–mesenchymal transition
  116. Visfatin is negatively associated with coronary artery lesions in subjects with impaired fasting glucose
  117. Treatment and outcomes of mechanical complications of acute myocardial infarction during the Covid-19 era: A comparison with the pre-Covid-19 period. A systematic review and meta-analysis
  118. Neonatal stroke surveillance study protocol in the United Kingdom and Republic of Ireland
  119. Oncogenic role of TWF2 in human tumors: A pan-cancer analysis
  120. Mean corpuscular hemoglobin predicts the length of hospital stay independent of severity classification in patients with acute pancreatitis
  121. Association of gallstone and polymorphisms of UGT1A1*27 and UGT1A1*28 in patients with hepatitis B virus-related liver failure
  122. TGF-β1 upregulates Sar1a expression and induces procollagen-I secretion in hypertrophic scarring fibroblasts
  123. Antisense lncRNA PCNA-AS1 promotes esophageal squamous cell carcinoma progression through the miR-2467-3p/PCNA axis
  124. NK-cell dysfunction of acute myeloid leukemia in relation to the renin–angiotensin system and neurotransmitter genes
  125. The effect of dilution with glucose and prolonged injection time on dexamethasone-induced perineal irritation – A randomized controlled trial
  126. miR-146-5p restrains calcification of vascular smooth muscle cells by suppressing TRAF6
  127. Role of lncRNA MIAT/miR-361-3p/CCAR2 in prostate cancer cells
  128. lncRNA NORAD promotes lung cancer progression by competitively binding to miR-28-3p with E2F2
  129. Noninvasive diagnosis of AIH/PBC overlap syndrome based on prediction models
  130. lncRNA FAM230B is highly expressed in colorectal cancer and suppresses the maturation of miR-1182 to increase cell proliferation
  131. circ-LIMK1 regulates cisplatin resistance in lung adenocarcinoma by targeting miR-512-5p/HMGA1 axis
  132. LncRNA SNHG3 promoted cell proliferation, migration, and metastasis of esophageal squamous cell carcinoma via regulating miR-151a-3p/PFN2 axis
  133. Risk perception and affective state on work exhaustion in obstetrics during the COVID-19 pandemic
  134. lncRNA-AC130710/miR-129-5p/mGluR1 axis promote migration and invasion by activating PKCα-MAPK signal pathway in melanoma
  135. SNRPB promotes cell cycle progression in thyroid carcinoma via inhibiting p53
  136. Xylooligosaccharides and aerobic training regulate metabolism and behavior in rats with streptozotocin-induced type 1 diabetes
  137. Serpin family A member 1 is an oncogene in glioma and its translation is enhanced by NAD(P)H quinone dehydrogenase 1 through RNA-binding activity
  138. Silencing of CPSF7 inhibits the proliferation, migration, and invasion of lung adenocarcinoma cells by blocking the AKT/mTOR signaling pathway
  139. Ultrasound-guided lumbar plexus block versus transversus abdominis plane block for analgesia in children with hip dislocation: A double-blind, randomized trial
  140. Relationship of plasma MBP and 8-oxo-dG with brain damage in preterm
  141. Identification of a novel necroptosis-associated miRNA signature for predicting the prognosis in head and neck squamous cell carcinoma
  142. Delayed femoral vein ligation reduces operative time and blood loss during hip disarticulation in patients with extremity tumors
  143. The expression of ASAP3 and NOTCH3 and the clinicopathological characteristics of adult glioma patients
  144. Longitudinal analysis of factors related to Helicobacter pylori infection in Chinese adults
  145. HOXA10 enhances cell proliferation and suppresses apoptosis in esophageal cancer via activating p38/ERK signaling pathway
  146. Meta-analysis of early-life antibiotic use and allergic rhinitis
  147. Marital status and its correlation with age, race, and gender in prognosis of tonsil squamous cell carcinomas
  148. HPV16 E6E7 up-regulates KIF2A expression by activating JNK/c-Jun signal, is beneficial to migration and invasion of cervical cancer cells
  149. Amino acid profiles in the tissue and serum of patients with liver cancer
  150. Pain in critically ill COVID-19 patients: An Italian retrospective study
  151. Immunohistochemical distribution of Bcl-2 and p53 apoptotic markers in acetamiprid-induced nephrotoxicity
  152. Estradiol pretreatment in GnRH antagonist protocol for IVF/ICSI treatment
  153. Long non-coding RNAs LINC00689 inhibits the apoptosis of human nucleus pulposus cells via miR-3127-5p/ATG7 axis-mediated autophagy
  154. The relationship between oxygen therapy, drug therapy, and COVID-19 mortality
  155. Monitoring hypertensive disorders in pregnancy to prevent preeclampsia in pregnant women of advanced maternal age: Trial mimicking with retrospective data
  156. SETD1A promotes the proliferation and glycolysis of nasopharyngeal carcinoma cells by activating the PI3K/Akt pathway
  157. The role of Shunaoxin pills in the treatment of chronic cerebral hypoperfusion and its main pharmacodynamic components
  158. TET3 governs malignant behaviors and unfavorable prognosis of esophageal squamous cell carcinoma by activating the PI3K/AKT/GSK3β/β-catenin pathway
  159. Associations between morphokinetic parameters of temporary-arrest embryos and the clinical prognosis in FET cycles
  160. Long noncoding RNA WT1-AS regulates trophoblast proliferation, migration, and invasion via the microRNA-186-5p/CADM2 axis
  161. The incidence of bronchiectasis in chronic obstructive pulmonary disease
  162. Integrated bioinformatics analysis shows integrin alpha 3 is a prognostic biomarker for pancreatic cancer
  163. Inhibition of miR-21 improves pulmonary vascular responses in bronchopulmonary dysplasia by targeting the DDAH1/ADMA/NO pathway
  164. Comparison of hospitalized patients with severe pneumonia caused by COVID-19 and influenza A (H7N9 and H1N1): A retrospective study from a designated hospital
  165. lncRNA ZFAS1 promotes intervertebral disc degeneration by upregulating AAK1
  166. Pathological characteristics of liver injury induced by N,N-dimethylformamide: From humans to animal models
  167. lncRNA ELFN1-AS1 enhances the progression of colon cancer by targeting miR-4270 to upregulate AURKB
  168. DARS-AS1 modulates cell proliferation and migration of gastric cancer cells by regulating miR-330-3p/NAT10 axis
  169. Dezocine inhibits cell proliferation, migration, and invasion by targeting CRABP2 in ovarian cancer
  170. MGST1 alleviates the oxidative stress of trophoblast cells induced by hypoxia/reoxygenation and promotes cell proliferation, migration, and invasion by activating the PI3K/AKT/mTOR pathway
  171. Bifidobacterium lactis Probio-M8 ameliorated the symptoms of type 2 diabetes mellitus mice by changing ileum FXR-CYP7A1
  172. circRNA DENND1B inhibits tumorigenicity of clear cell renal cell carcinoma via miR-122-5p/TIMP2 axis
  173. EphA3 targeted by miR-3666 contributes to melanoma malignancy via activating ERK1/2 and p38 MAPK pathways
  174. Pacemakers and methylprednisolone pulse therapy in immune-related myocarditis concomitant with complete heart block
  175. miRNA-130a-3p targets sphingosine-1-phosphate receptor 1 to activate the microglial and astrocytes and to promote neural injury under the high glucose condition
  176. Review Articles
  177. Current management of cancer pain in Italy: Expert opinion paper
  178. Hearing loss and brain disorders: A review of multiple pathologies
  179. The rationale for using low-molecular weight heparin in the therapy of symptomatic COVID-19 patients
  180. Amyotrophic lateral sclerosis and delayed onset muscle soreness in light of the impaired blink and stretch reflexes – watch out for Piezo2
  181. Interleukin-35 in autoimmune dermatoses: Current concepts
  182. Recent discoveries in microbiota dysbiosis, cholangiocytic factors, and models for studying the pathogenesis of primary sclerosing cholangitis
  183. Advantages of ketamine in pediatric anesthesia
  184. Congenital adrenal hyperplasia. Role of dentist in early diagnosis
  185. Migraine management: Non-pharmacological points for patients and health care professionals
  186. Atherogenic index of plasma and coronary artery disease: A systematic review
  187. Physiological and modulatory role of thioredoxins in the cellular function
  188. Case Reports
  189. Intrauterine Bakri balloon tamponade plus cervical cerclage for the prevention and treatment of postpartum haemorrhage in late pregnancy complicated with acute aortic dissection: Case series
  190. A case of successful pembrolizumab monotherapy in a patient with advanced lung adenocarcinoma: Use of multiple biomarkers in combination for clinical practice
  191. Unusual neurological manifestations of bilateral medial medullary infarction: A case report
  192. Atypical symptoms of malignant hyperthermia: A rare causative mutation in the RYR1 gene
  193. A case report of dermatomyositis with the missed diagnosis of non-small cell lung cancer and concurrence of pulmonary tuberculosis
  194. A rare case of endometrial polyp complicated with uterine inversion: A case report and clinical management
  195. Spontaneous rupturing of splenic artery aneurysm: Another reason for fatal syncope and shock (Case report and literature review)
  196. Fungal infection mimicking COVID-19 infection – A case report
  197. Concurrent aspergillosis and cystic pulmonary metastases in a patient with tongue squamous cell carcinoma
  198. Paraganglioma-induced inverted takotsubo-like cardiomyopathy leading to cardiogenic shock successfully treated with extracorporeal membrane oxygenation
  199. Lineage switch from lymphoma to myeloid neoplasms: First case series from a single institution
  200. Trismus during tracheal extubation as a complication of general anaesthesia – A case report
  201. Simultaneous treatment of a pubovesical fistula and lymph node metastasis secondary to multimodal treatment for prostate cancer: Case report and review of the literature
  202. Two case reports of skin vasculitis following the COVID-19 immunization
  203. Ureteroiliac fistula after oncological surgery: Case report and review of the literature
  204. Synchronous triple primary malignant tumours in the bladder, prostate, and lung harbouring TP53 and MEK1 mutations accompanied with severe cardiovascular diseases: A case report
  205. Huge mucinous cystic neoplasms with adhesion to the left colon: A case report and literature review
  206. Commentary
  207. Commentary on “Clinicopathological features of programmed cell death-ligand 1 expression in patients with oral squamous cell carcinoma”
  208. Rapid Communication
  209. COVID-19 fear, post-traumatic stress, growth, and the role of resilience
  210. Erratum
  211. Erratum to “Tollip promotes hepatocellular carcinoma progression via PI3K/AKT pathway”
  212. Erratum to “Effect of femoral head necrosis cystic area on femoral head collapse and stress distribution in femoral head: A clinical and finite element study”
  213. Erratum to “lncRNA NORAD promotes lung cancer progression by competitively binding to miR-28-3p with E2F2”
  214. Retraction
  215. Expression and role of ABIN1 in sepsis: In vitro and in vivo studies
  216. Retraction to “miR-519d downregulates LEP expression to inhibit preeclampsia development”
  217. Special Issue Computational Intelligence Methodologies Meets Recurrent Cancers - Part II
  218. Usefulness of close surveillance for rectal cancer patients after neoadjuvant chemoradiotherapy
https://doi.org/10.1515/med-2022-0446
Keywords for this article
lncRNA NORAD; acute lung injury; HPMECs; miR-30c-5p; glucose metabolism; LDHA
Creative Commons
BY 4.0

Articles in the same Issue

  1. Research Articles
  2. AMBRA1 attenuates the proliferation of uveal melanoma cells
  3. A ceRNA network mediated by LINC00475 in papillary thyroid carcinoma
  4. Differences in complications between hepatitis B-related cirrhosis and alcohol-related cirrhosis
  5. Effect of gestational diabetes mellitus on lipid profile: A systematic review and meta-analysis
  6. Long noncoding RNA NR2F1-AS1 stimulates the tumorigenic behavior of non-small cell lung cancer cells by sponging miR-363-3p to increase SOX4
  7. Promising novel biomarkers and candidate small-molecule drugs for lung adenocarcinoma: Evidence from bioinformatics analysis of high-throughput data
  8. Plasmapheresis: Is it a potential alternative treatment for chronic urticaria?
  9. The biomarkers of key miRNAs and gene targets associated with extranodal NK/T-cell lymphoma
  10. Gene signature to predict prognostic survival of hepatocellular carcinoma
  11. Effects of miRNA-199a-5p on cell proliferation and apoptosis of uterine leiomyoma by targeting MED12
  12. Does diabetes affect paraneoplastic thrombocytosis in colorectal cancer?
  13. Is there any effect on imprinted genes H19, PEG3, and SNRPN during AOA?
  14. Leptin and PCSK9 concentrations are associated with vascular endothelial cytokines in patients with stable coronary heart disease
  15. Pericentric inversion of chromosome 6 and male fertility problems
  16. Staple line reinforcement with nebulized cyanoacrylate glue in laparoscopic sleeve gastrectomy: A propensity score-matched study
  17. Retrospective analysis of crescent score in clinical prognosis of IgA nephropathy
  18. Expression of DNM3 is associated with good outcome in colorectal cancer
  19. Activation of SphK2 contributes to adipocyte-induced EOC cell proliferation
  20. CRRT influences PICCO measurements in febrile critically ill patients
  21. SLCO4A1-AS1 mediates pancreatic cancer development via miR-4673/KIF21B axis
  22. lncRNA ACTA2-AS1 inhibits malignant phenotypes of gastric cancer cells
  23. circ_AKT3 knockdown suppresses cisplatin resistance in gastric cancer
  24. Prognostic value of nicotinamide N-methyltransferase in human cancers: Evidence from a meta-analysis and database validation
  25. GPC2 deficiency inhibits cell growth and metastasis in colon adenocarcinoma
  26. A pan-cancer analysis of the oncogenic role of Holliday junction recognition protein in human tumors
  27. Radiation increases COL1A1, COL3A1, and COL1A2 expression in breast cancer
  28. Association between preventable risk factors and metabolic syndrome
  29. miR-29c-5p knockdown reduces inflammation and blood–brain barrier disruption by upregulating LRP6
  30. Cardiac contractility modulation ameliorates myocardial metabolic remodeling in a rabbit model of chronic heart failure through activation of AMPK and PPAR-α pathway
  31. Quercitrin protects human bronchial epithelial cells from oxidative damage
  32. Smurf2 suppresses the metastasis of hepatocellular carcinoma via ubiquitin degradation of Smad2
  33. circRNA_0001679/miR-338-3p/DUSP16 axis aggravates acute lung injury
  34. Sonoclot’s usefulness in prediction of cardiopulmonary arrest prognosis: A proof of concept study
  35. Four drug metabolism-related subgroups of pancreatic adenocarcinoma in prognosis, immune infiltration, and gene mutation
  36. Decreased expression of miR-195 mediated by hypermethylation promotes osteosarcoma
  37. LMO3 promotes proliferation and metastasis of papillary thyroid carcinoma cells by regulating LIMK1-mediated cofilin and the β-catenin pathway
  38. Cx43 upregulation in HUVECs under stretch via TGF-β1 and cytoskeletal network
  39. Evaluation of menstrual irregularities after COVID-19 vaccination: Results of the MECOVAC survey
  40. Histopathologic findings on removed stomach after sleeve gastrectomy. Do they influence the outcome?
  41. Analysis of the expression and prognostic value of MT1-MMP, β1-integrin and YAP1 in glioma
  42. Optimal diagnosis of the skin cancer using a hybrid deep neural network and grasshopper optimization algorithm
  43. miR-223-3p alleviates TGF-β-induced epithelial-mesenchymal transition and extracellular matrix deposition by targeting SP3 in endometrial epithelial cells
  44. Clinical value of SIRT1 as a prognostic biomarker in esophageal squamous cell carcinoma, a systematic meta-analysis
  45. circ_0020123 promotes cell proliferation and migration in lung adenocarcinoma via PDZD8
  46. miR-22-5p regulates the self-renewal of spermatogonial stem cells by targeting EZH2
  47. hsa-miR-340-5p inhibits epithelial–mesenchymal transition in endometriosis by targeting MAP3K2 and inactivating MAPK/ERK signaling
  48. circ_0085296 inhibits the biological functions of trophoblast cells to promote the progression of preeclampsia via the miR-942-5p/THBS2 network
  49. TCD hemodynamics findings in the subacute phase of anterior circulation stroke patients treated with mechanical thrombectomy
  50. Development of a risk-stratification scoring system for predicting risk of breast cancer based on non-alcoholic fatty liver disease, non-alcoholic fatty pancreas disease, and uric acid
  51. Tollip promotes hepatocellular carcinoma progression via PI3K/AKT pathway
  52. circ_0062491 alleviates periodontitis via the miR-142-5p/IGF1 axis
  53. Human amniotic fluid as a source of stem cells
  54. lncRNA NONRATT013819.2 promotes transforming growth factor-β1-induced myofibroblastic transition of hepatic stellate cells by miR24-3p/lox
  55. NORAD modulates miR-30c-5p-LDHA to protect lung endothelial cells damage
  56. Idiopathic pulmonary fibrosis telemedicine management during COVID-19 outbreak
  57. Risk factors for adverse drug reactions associated with clopidogrel therapy
  58. Serum zinc associated with immunity and inflammatory markers in Covid-19
  59. The relationship between night shift work and breast cancer incidence: A systematic review and meta-analysis of observational studies
  60. LncRNA expression in idiopathic achalasia: New insight and preliminary exploration into pathogenesis
  61. Notoginsenoside R1 alleviates spinal cord injury through the miR-301a/KLF7 axis to activate Wnt/β-catenin pathway
  62. Moscatilin suppresses the inflammation from macrophages and T cells
  63. Zoledronate promotes ECM degradation and apoptosis via Wnt/β-catenin
  64. Epithelial-mesenchymal transition-related genes in coronary artery disease
  65. The effect evaluation of traditional vaginal surgery and transvaginal mesh surgery for severe pelvic organ prolapse: 5 years follow-up
  66. Repeated partial splenic artery embolization for hypersplenism improves platelet count
  67. Low expression of miR-27b in serum exosomes of non-small cell lung cancer facilitates its progression by affecting EGFR
  68. Exosomal hsa_circ_0000519 modulates the NSCLC cell growth and metastasis via miR-1258/RHOV axis
  69. miR-455-5p enhances 5-fluorouracil sensitivity in colorectal cancer cells by targeting PIK3R1 and DEPDC1
  70. The effect of tranexamic acid on the reduction of intraoperative and postoperative blood loss and thromboembolic risk in patients with hip fracture
  71. Isocitrate dehydrogenase 1 mutation in cholangiocarcinoma impairs tumor progression by sensitizing cells to ferroptosis
  72. Artemisinin protects against cerebral ischemia and reperfusion injury via inhibiting the NF-κB pathway
  73. A 16-gene signature associated with homologous recombination deficiency for prognosis prediction in patients with triple-negative breast cancer
  74. Lidocaine ameliorates chronic constriction injury-induced neuropathic pain through regulating M1/M2 microglia polarization
  75. MicroRNA 322-5p reduced neuronal inflammation via the TLR4/TRAF6/NF-κB axis in a rat epilepsy model
  76. miR-1273h-5p suppresses CXCL12 expression and inhibits gastric cancer cell invasion and metastasis
  77. Clinical characteristics of pneumonia patients of long course of illness infected with SARS-CoV-2
  78. circRNF20 aggravates the malignancy of retinoblastoma depending on the regulation of miR-132-3p/PAX6 axis
  79. Linezolid for resistant Gram-positive bacterial infections in children under 12 years: A meta-analysis
  80. Rack1 regulates pro-inflammatory cytokines by NF-κB in diabetic nephropathy
  81. Comprehensive analysis of molecular mechanism and a novel prognostic signature based on small nuclear RNA biomarkers in gastric cancer patients
  82. Smog and risk of maternal and fetal birth outcomes: A retrospective study in Baoding, China
  83. Let-7i-3p inhibits the cell cycle, proliferation, invasion, and migration of colorectal cancer cells via downregulating CCND1
  84. β2-Adrenergic receptor expression in subchondral bone of patients with varus knee osteoarthritis
  85. Possible impact of COVID-19 pandemic and lockdown on suicide behavior among patients in Southeast Serbia
  86. In vitro antimicrobial activity of ozonated oil in liposome eyedrop against multidrug-resistant bacteria
  87. Potential biomarkers for inflammatory response in acute lung injury
  88. A low serum uric acid concentration predicts a poor prognosis in adult patients with candidemia
  89. Antitumor activity of recombinant oncolytic vaccinia virus with human IL2
  90. ALKBH5 inhibits TNF-α-induced apoptosis of HUVECs through Bcl-2 pathway
  91. Risk prediction of cardiovascular disease using machine learning classifiers
  92. Value of ultrasonography parameters in diagnosing polycystic ovary syndrome
  93. Bioinformatics analysis reveals three key genes and four survival genes associated with youth-onset NSCLC
  94. Identification of autophagy-related biomarkers in patients with pulmonary arterial hypertension based on bioinformatics analysis
  95. Protective effects of glaucocalyxin A on the airway of asthmatic mice
  96. Overexpression of miR-100-5p inhibits papillary thyroid cancer progression via targeting FZD8
  97. Bioinformatics-based analysis of SUMOylation-related genes in hepatocellular carcinoma reveals a role of upregulated SAE1 in promoting cell proliferation
  98. Effectiveness and clinical benefits of new anti-diabetic drugs: A real life experience
  99. Identification of osteoporosis based on gene biomarkers using support vector machine
  100. Tanshinone IIA reverses oxaliplatin resistance in colorectal cancer through microRNA-30b-5p/AVEN axis
  101. miR-212-5p inhibits nasopharyngeal carcinoma progression by targeting METTL3
  102. Association of ST-T changes with all-cause mortality among patients with peripheral T-cell lymphomas
  103. LINC00665/miRNAs axis-mediated collagen type XI alpha 1 correlates with immune infiltration and malignant phenotypes in lung adenocarcinoma
  104. The perinatal factors that influence the excretion of fecal calprotectin in premature-born children
  105. Effect of femoral head necrosis cystic area on femoral head collapse and stress distribution in femoral head: A clinical and finite element study
  106. Does the use of 3D-printed cones give a chance to postpone the use of megaprostheses in patients with large bone defects in the knee joint?
  107. lncRNA HAGLR modulates myocardial ischemia–reperfusion injury in mice through regulating miR-133a-3p/MAPK1 axis
  108. Protective effect of ghrelin on intestinal I/R injury in rats
  109. In vivo knee kinematics of an innovative prosthesis design
  110. Relationship between the height of fibular head and the incidence and severity of knee osteoarthritis
  111. lncRNA WT1-AS attenuates hypoxia/ischemia-induced neuronal injury during cerebral ischemic stroke via miR-186-5p/XIAP axis
  112. Correlation of cardiac troponin T and APACHE III score with all-cause in-hospital mortality in critically ill patients with acute pulmonary embolism
  113. LncRNA LINC01857 reduces metastasis and angiogenesis in breast cancer cells via regulating miR-2052/CENPQ axis
  114. Endothelial cell-specific molecule 1 (ESM1) promoted by transcription factor SPI1 acts as an oncogene to modulate the malignant phenotype of endometrial cancer
  115. SELENBP1 inhibits progression of colorectal cancer by suppressing epithelial–mesenchymal transition
  116. Visfatin is negatively associated with coronary artery lesions in subjects with impaired fasting glucose
  117. Treatment and outcomes of mechanical complications of acute myocardial infarction during the Covid-19 era: A comparison with the pre-Covid-19 period. A systematic review and meta-analysis
  118. Neonatal stroke surveillance study protocol in the United Kingdom and Republic of Ireland
  119. Oncogenic role of TWF2 in human tumors: A pan-cancer analysis
  120. Mean corpuscular hemoglobin predicts the length of hospital stay independent of severity classification in patients with acute pancreatitis
  121. Association of gallstone and polymorphisms of UGT1A1*27 and UGT1A1*28 in patients with hepatitis B virus-related liver failure
  122. TGF-β1 upregulates Sar1a expression and induces procollagen-I secretion in hypertrophic scarring fibroblasts
  123. Antisense lncRNA PCNA-AS1 promotes esophageal squamous cell carcinoma progression through the miR-2467-3p/PCNA axis
  124. NK-cell dysfunction of acute myeloid leukemia in relation to the renin–angiotensin system and neurotransmitter genes
  125. The effect of dilution with glucose and prolonged injection time on dexamethasone-induced perineal irritation – A randomized controlled trial
  126. miR-146-5p restrains calcification of vascular smooth muscle cells by suppressing TRAF6
  127. Role of lncRNA MIAT/miR-361-3p/CCAR2 in prostate cancer cells
  128. lncRNA NORAD promotes lung cancer progression by competitively binding to miR-28-3p with E2F2
  129. Noninvasive diagnosis of AIH/PBC overlap syndrome based on prediction models
  130. lncRNA FAM230B is highly expressed in colorectal cancer and suppresses the maturation of miR-1182 to increase cell proliferation
  131. circ-LIMK1 regulates cisplatin resistance in lung adenocarcinoma by targeting miR-512-5p/HMGA1 axis
  132. LncRNA SNHG3 promoted cell proliferation, migration, and metastasis of esophageal squamous cell carcinoma via regulating miR-151a-3p/PFN2 axis
  133. Risk perception and affective state on work exhaustion in obstetrics during the COVID-19 pandemic
  134. lncRNA-AC130710/miR-129-5p/mGluR1 axis promote migration and invasion by activating PKCα-MAPK signal pathway in melanoma
  135. SNRPB promotes cell cycle progression in thyroid carcinoma via inhibiting p53
  136. Xylooligosaccharides and aerobic training regulate metabolism and behavior in rats with streptozotocin-induced type 1 diabetes
  137. Serpin family A member 1 is an oncogene in glioma and its translation is enhanced by NAD(P)H quinone dehydrogenase 1 through RNA-binding activity
  138. Silencing of CPSF7 inhibits the proliferation, migration, and invasion of lung adenocarcinoma cells by blocking the AKT/mTOR signaling pathway
  139. Ultrasound-guided lumbar plexus block versus transversus abdominis plane block for analgesia in children with hip dislocation: A double-blind, randomized trial
  140. Relationship of plasma MBP and 8-oxo-dG with brain damage in preterm
  141. Identification of a novel necroptosis-associated miRNA signature for predicting the prognosis in head and neck squamous cell carcinoma
  142. Delayed femoral vein ligation reduces operative time and blood loss during hip disarticulation in patients with extremity tumors
  143. The expression of ASAP3 and NOTCH3 and the clinicopathological characteristics of adult glioma patients
  144. Longitudinal analysis of factors related to Helicobacter pylori infection in Chinese adults
  145. HOXA10 enhances cell proliferation and suppresses apoptosis in esophageal cancer via activating p38/ERK signaling pathway
  146. Meta-analysis of early-life antibiotic use and allergic rhinitis
  147. Marital status and its correlation with age, race, and gender in prognosis of tonsil squamous cell carcinomas
  148. HPV16 E6E7 up-regulates KIF2A expression by activating JNK/c-Jun signal, is beneficial to migration and invasion of cervical cancer cells
  149. Amino acid profiles in the tissue and serum of patients with liver cancer
  150. Pain in critically ill COVID-19 patients: An Italian retrospective study
  151. Immunohistochemical distribution of Bcl-2 and p53 apoptotic markers in acetamiprid-induced nephrotoxicity
  152. Estradiol pretreatment in GnRH antagonist protocol for IVF/ICSI treatment
  153. Long non-coding RNAs LINC00689 inhibits the apoptosis of human nucleus pulposus cells via miR-3127-5p/ATG7 axis-mediated autophagy
  154. The relationship between oxygen therapy, drug therapy, and COVID-19 mortality
  155. Monitoring hypertensive disorders in pregnancy to prevent preeclampsia in pregnant women of advanced maternal age: Trial mimicking with retrospective data
  156. SETD1A promotes the proliferation and glycolysis of nasopharyngeal carcinoma cells by activating the PI3K/Akt pathway
  157. The role of Shunaoxin pills in the treatment of chronic cerebral hypoperfusion and its main pharmacodynamic components
  158. TET3 governs malignant behaviors and unfavorable prognosis of esophageal squamous cell carcinoma by activating the PI3K/AKT/GSK3β/β-catenin pathway
  159. Associations between morphokinetic parameters of temporary-arrest embryos and the clinical prognosis in FET cycles
  160. Long noncoding RNA WT1-AS regulates trophoblast proliferation, migration, and invasion via the microRNA-186-5p/CADM2 axis
  161. The incidence of bronchiectasis in chronic obstructive pulmonary disease
  162. Integrated bioinformatics analysis shows integrin alpha 3 is a prognostic biomarker for pancreatic cancer
  163. Inhibition of miR-21 improves pulmonary vascular responses in bronchopulmonary dysplasia by targeting the DDAH1/ADMA/NO pathway
  164. Comparison of hospitalized patients with severe pneumonia caused by COVID-19 and influenza A (H7N9 and H1N1): A retrospective study from a designated hospital
  165. lncRNA ZFAS1 promotes intervertebral disc degeneration by upregulating AAK1
  166. Pathological characteristics of liver injury induced by N,N-dimethylformamide: From humans to animal models
  167. lncRNA ELFN1-AS1 enhances the progression of colon cancer by targeting miR-4270 to upregulate AURKB
  168. DARS-AS1 modulates cell proliferation and migration of gastric cancer cells by regulating miR-330-3p/NAT10 axis
  169. Dezocine inhibits cell proliferation, migration, and invasion by targeting CRABP2 in ovarian cancer
  170. MGST1 alleviates the oxidative stress of trophoblast cells induced by hypoxia/reoxygenation and promotes cell proliferation, migration, and invasion by activating the PI3K/AKT/mTOR pathway
  171. Bifidobacterium lactis Probio-M8 ameliorated the symptoms of type 2 diabetes mellitus mice by changing ileum FXR-CYP7A1
  172. circRNA DENND1B inhibits tumorigenicity of clear cell renal cell carcinoma via miR-122-5p/TIMP2 axis
  173. EphA3 targeted by miR-3666 contributes to melanoma malignancy via activating ERK1/2 and p38 MAPK pathways
  174. Pacemakers and methylprednisolone pulse therapy in immune-related myocarditis concomitant with complete heart block
  175. miRNA-130a-3p targets sphingosine-1-phosphate receptor 1 to activate the microglial and astrocytes and to promote neural injury under the high glucose condition
  176. Review Articles
  177. Current management of cancer pain in Italy: Expert opinion paper
  178. Hearing loss and brain disorders: A review of multiple pathologies
  179. The rationale for using low-molecular weight heparin in the therapy of symptomatic COVID-19 patients
  180. Amyotrophic lateral sclerosis and delayed onset muscle soreness in light of the impaired blink and stretch reflexes – watch out for Piezo2
  181. Interleukin-35 in autoimmune dermatoses: Current concepts
  182. Recent discoveries in microbiota dysbiosis, cholangiocytic factors, and models for studying the pathogenesis of primary sclerosing cholangitis
  183. Advantages of ketamine in pediatric anesthesia
  184. Congenital adrenal hyperplasia. Role of dentist in early diagnosis
  185. Migraine management: Non-pharmacological points for patients and health care professionals
  186. Atherogenic index of plasma and coronary artery disease: A systematic review
  187. Physiological and modulatory role of thioredoxins in the cellular function
  188. Case Reports
  189. Intrauterine Bakri balloon tamponade plus cervical cerclage for the prevention and treatment of postpartum haemorrhage in late pregnancy complicated with acute aortic dissection: Case series
  190. A case of successful pembrolizumab monotherapy in a patient with advanced lung adenocarcinoma: Use of multiple biomarkers in combination for clinical practice
  191. Unusual neurological manifestations of bilateral medial medullary infarction: A case report
  192. Atypical symptoms of malignant hyperthermia: A rare causative mutation in the RYR1 gene
  193. A case report of dermatomyositis with the missed diagnosis of non-small cell lung cancer and concurrence of pulmonary tuberculosis
  194. A rare case of endometrial polyp complicated with uterine inversion: A case report and clinical management
  195. Spontaneous rupturing of splenic artery aneurysm: Another reason for fatal syncope and shock (Case report and literature review)
  196. Fungal infection mimicking COVID-19 infection – A case report
  197. Concurrent aspergillosis and cystic pulmonary metastases in a patient with tongue squamous cell carcinoma
  198. Paraganglioma-induced inverted takotsubo-like cardiomyopathy leading to cardiogenic shock successfully treated with extracorporeal membrane oxygenation
  199. Lineage switch from lymphoma to myeloid neoplasms: First case series from a single institution
  200. Trismus during tracheal extubation as a complication of general anaesthesia – A case report
  201. Simultaneous treatment of a pubovesical fistula and lymph node metastasis secondary to multimodal treatment for prostate cancer: Case report and review of the literature
  202. Two case reports of skin vasculitis following the COVID-19 immunization
  203. Ureteroiliac fistula after oncological surgery: Case report and review of the literature
  204. Synchronous triple primary malignant tumours in the bladder, prostate, and lung harbouring TP53 and MEK1 mutations accompanied with severe cardiovascular diseases: A case report
  205. Huge mucinous cystic neoplasms with adhesion to the left colon: A case report and literature review
  206. Commentary
  207. Commentary on “Clinicopathological features of programmed cell death-ligand 1 expression in patients with oral squamous cell carcinoma”
  208. Rapid Communication
  209. COVID-19 fear, post-traumatic stress, growth, and the role of resilience
  210. Erratum
  211. Erratum to “Tollip promotes hepatocellular carcinoma progression via PI3K/AKT pathway”
  212. Erratum to “Effect of femoral head necrosis cystic area on femoral head collapse and stress distribution in femoral head: A clinical and finite element study”
  213. Erratum to “lncRNA NORAD promotes lung cancer progression by competitively binding to miR-28-3p with E2F2”
  214. Retraction
  215. Expression and role of ABIN1 in sepsis: In vitro and in vivo studies
  216. Retraction to “miR-519d downregulates LEP expression to inhibit preeclampsia development”
  217. Special Issue Computational Intelligence Methodologies Meets Recurrent Cancers - Part II
  218. Usefulness of close surveillance for rectal cancer patients after neoadjuvant chemoradiotherapy
Sign up now to receive a 20% welcome discount
Institutional Access
How does access work?
Winner of the OpenAthens UX Award 2026
Winner of the OpenAthens UX Award 2026
Have an idea on how to improve our website?
Please write us.
© 2026 De Gruyter Brill
Downloaded on 31.3.2026 from https://www.degruyterbrill.com/document/doi/10.1515/med-2022-0446/html
Scroll to top button