Kinetic characterization of rat brain acetylcholinesterase modulated by lead and cartap: the ameliorative effect of Citrus limon fruit juice
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Nitika Singh
Abstract
Objectives
Human exposure to heavy metals and pesticides is a worldwide major health problem. These environmental pollutants have been considered as the most neurotoxic agents and responsible to causing neurological toxicity. Plant-based therapeutic supplement may be used in the event of toxicity. Citrus limon contains several useful bioactive ingredients including flavonoids, dietary fiber, carotenoids, vitamins, pectin, minerals, and essential oils, which are responsible for its therapeutic potential. In the present investigation, we have studied the toxicity of heavy metals such as lead (Pb) and a carbamate pesticide such as cartap (Cp) on rat brain acetylcholinesterase (AChE).
Methods
The chemical characterization of C. limon involved determination of total antioxidants and 2,2-diphenyl-1-picryl-hydrazyl-hydrate free radical scavenging activity using known methods. The AChE activity and its kinetic characterization were performed by assaying the enzyme activity at varying substrate concentrations, pH, temperature, and time of reaction. Its different kinetic parameters such as K i , V max, K m , K cat, and K cat/K m were determined by using standard procedures. The amelioration potential of the extract was evaluated on the neurotransmission system of rat brain AChE treated with Pb, Cp, and their combination (Pb–Cp), considering their 50% inhibitory concentration (IC50) values.
Results
The optimal activity of rat brain AChE was recorded at 25 µg of protein, pH of 7.4, substrate concentration [S] of 0.5 mM, and temperature of 37.4–40°C. The enzyme was stable for 10 min when incubated at 37.4 °C in vitro. The enzyme displayed 70% of its activity remaining even after 160 min of incubation in this condition. It may be stable up to 1 month when stored at −20°C. The IC50 values for Pb, Cp, and Pb–Cp were found to be 75, 2.9, and 5 mM, respectively. Pb, Cp, and Pb–Cp inhibited the activity of rat brain AChE in the noncompetitive, mixed, and uncompetitive manners, respectively, with their respective K i values to be 675, 2.37, and 22.72 mM.
Conclusions
The results indicated that the Pb and Cp were able to cause significant alterations in the level and properties of AChE. However, the introduction of lemon juice on Pb- and Cp-treated AChE indicated protection of its activity from their adverse effects. The results may be useful in prospective therapeutic applications of lemon juice or as a food supplement to protect mammalian systems from adverse effects of these toxicants.
Funding source: University Grant Commission
Funding source: UPCST-Lucknow
Acknowledgments
The authors acknowledge UGC-SAP and DST-FIST for support to Department of Biochemistry, University of Allahabad, Allahabad, India.
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Research funding: NS, PT, and BS are grateful to the University Grant Commission, New Delhi, and UPCST-Lucknow for providing financial assistance in the forms of a Research Fellowship to NS and a research grant to BS.
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Author contributions: NS and PT performed the experiments, wrote the article, prepared and assembled the figures and tables; BS planned the experiments, reviewed and critically organized the article.
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Competing interests: The authors state no conflict of interest.
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Ethical approval: The approval was received from the Institutional Animal Ethics Committee (IAEC).
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© 2020 Walter de Gruyter GmbH, Berlin/Boston
Artikel in diesem Heft
- Frontmatter
- Reviews
- An insight on safety, efficacy, and molecular docking study reports of N-acetylcysteine and its compound formulations
- Public health approach of Unani medicine to cope and stay safe in hot environmental conditions
- Original Articles
- Kinetic characterization of rat brain acetylcholinesterase modulated by lead and cartap: the ameliorative effect of Citrus limon fruit juice
- Attenuation of potassium dichromate and sodium arsenite toxicities by methanol extract of Rauvolfia vomitoria in mice
- Chronic exposure of industrial grade calcium carbide and ethylene glycol alter histological architecture of systemic organs by disrupting redox balance in rat
- Biochemical and histopathological effects of low dose vanadium in the healing of acetic acid-induced colitis in male wistar rats
- Glycine and L-Arginine supplementation ameliorates gastro-duodenal toxicity in a rat model of NSAID (Diclofenac)-gastroenteropathy via inhibition of oxidative stress
- Cocos nucifera L. oil alleviates lead acetate-induced reproductive toxicity in sexually-matured male Wistar rats
- Intracellular signaling molecules of nerve tissue progenitors as pharmacological targets for treatment of ethanol-induced neurodegeneration
- Glatiramer acetate treatment inhibits inflammatory responses and improves survival in a mice model of cecal ligation and puncture-induced sepsis
- Ciprofloxacin-induced cutaneous adverse drug events: a systematic review of descriptive studies
- Aqueous extract of Adansonia digitata prevents Cadmium Chloride-induced testicular damage in Wistar Rats
- Exposure to prolonged unpredictable light impairs spatial memory via induction of oxidative stress and tumor necrosis factor-alpha in rats
- Tamarindus indica. Linn leaves ameliorates experimental induced heart failure in Wistar rats
Artikel in diesem Heft
- Frontmatter
- Reviews
- An insight on safety, efficacy, and molecular docking study reports of N-acetylcysteine and its compound formulations
- Public health approach of Unani medicine to cope and stay safe in hot environmental conditions
- Original Articles
- Kinetic characterization of rat brain acetylcholinesterase modulated by lead and cartap: the ameliorative effect of Citrus limon fruit juice
- Attenuation of potassium dichromate and sodium arsenite toxicities by methanol extract of Rauvolfia vomitoria in mice
- Chronic exposure of industrial grade calcium carbide and ethylene glycol alter histological architecture of systemic organs by disrupting redox balance in rat
- Biochemical and histopathological effects of low dose vanadium in the healing of acetic acid-induced colitis in male wistar rats
- Glycine and L-Arginine supplementation ameliorates gastro-duodenal toxicity in a rat model of NSAID (Diclofenac)-gastroenteropathy via inhibition of oxidative stress
- Cocos nucifera L. oil alleviates lead acetate-induced reproductive toxicity in sexually-matured male Wistar rats
- Intracellular signaling molecules of nerve tissue progenitors as pharmacological targets for treatment of ethanol-induced neurodegeneration
- Glatiramer acetate treatment inhibits inflammatory responses and improves survival in a mice model of cecal ligation and puncture-induced sepsis
- Ciprofloxacin-induced cutaneous adverse drug events: a systematic review of descriptive studies
- Aqueous extract of Adansonia digitata prevents Cadmium Chloride-induced testicular damage in Wistar Rats
- Exposure to prolonged unpredictable light impairs spatial memory via induction of oxidative stress and tumor necrosis factor-alpha in rats
- Tamarindus indica. Linn leaves ameliorates experimental induced heart failure in Wistar rats
