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Experimental validation of Vitex negundo leaves hydroalcoholic extract for neuroprotection in haloperidol induced parkinson’s disease in rat

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Abstract

Parkinsonism is a neurodegenerative disease, mainly imbalance in dopamine and acetylcholine neurotransimitter in mid brain, which manifestation of dysfunctions of extrapyramidal like akinesia, tremor, rigidity and catalepsy etc., even cognitive and memory loss. The current study is framed to evaluate the effect of Vitex negundo (VNL) leaf extract in Haloperidol induced PD in rats. In vitro studies of antioxidant capacity were checked via DPPH and NO assays and identified its Acetylcholinesterase (AChE) inhibitory activity. Secondly the In vivo study of anti-PD activity in Haloperidol induced in rats were evaluated by Rotarod, morris water maze (MWM), cooks pole climb (CPC), actophotometer, novel object recognition (NOR), and T-maze were utilized to assess extrapyramidal, cognitive and memory function. Thirdly, changes in biomarker level viz. (AChE), butyrylcholinesterase. (BChE) in hippocampus and cortex, reduced glutathione (GSH), malondialdehyde (MDA), total protein (TP), superoxide dismutase (SOD), catalase (CAT), and dopamine level in the whole brain were measured. Finally, histopathology of hippocampus and cortex was examined at 40x magnification to access restoring integrity and maintaining the architecture of neuronal cell in the treatment group compared to control group and L-DOPA as a standard treatment group. V. negundo showed potent antioxidant potency on scavenging of DPPH (IC50 84.81 μg/ml) and NO (IC50 133.20 μg/ml) and possess AChE inhibitory potency (IC50 114.35 μg/ml) by in vitro studies. The Rotarod, MWM, CPC, Actophotometer, NOR, T-maze demonstrated that Haloperidol group administration declines performance time, ELT, TL and decreases locomotion, cognitive and memory respectively. The treatment of VNL 100, 200, and 400 mg/kg p.o. significantly (p < 0.05 to p < 0.0001) reversed. Whole brain AChE, BChE, and MDA level were significantly raised and GSH, TP, SOD, CAT and Dopamine were significantly declined in Haloperidol treated group rats, especially V. negundo 400 mg/kg p.o. highly significantly ameliorate the Haloperidol group altered pathological changes through the restoration of the cholinergic function, enhancing the antioxidant defense and by increasing the dopaminergic function. The current study provides validation of V. negundo for its anti-PD activity and could be a valuable source for the treatment of PD in future.

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Data availability

The data are not publicly available due to their containing information that could compromise the privacy of research participants, but are available on request from the author.

Abbreviations

V. negundo :

V. negundo

VNL100 :

V. negundo 100 mg/kg Group

VNL200 :

V. negundo 200 mg/kg Group

VNL400 :

V. negundo 400 mg/kg Group

ACh :

Acetylcholine

AChE :

Acetylcholinesterase

AChI :

Acetylthiocholine iodide

BChI :

Butyrylcholines iodide

AD :

Alzheimer’s Disease

PD :

Parkinson’s Disease

APP :

Amyloid Precursor Protein

Aβ :

Amyloid Beta

BCh :

Butyrylcholine

BChE :

Butyrylcholinesterase

DPPH :

2,2-diphenyl-1-picrylhydrazyl

DTNB :

5,5′-dithio-bis(2-nitrobenzoic acid)

ELT :

Escape Latency Time

GSH :

Glutathione

H2O2 :

Hydrogen Peroxide

i.p:

Intraperitoneal

IC50 :

Inhibition Concentration

LPO :

Lipid Peroxidation

MDA :

Malondialdehyde

MWM :

Morris Water Maze

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Acknowledgements

Thanks to the Prof.(Dr) Sunil S Jalapure, Principal and Prof (Dr.) N A Khatib, HOD, Department of Pharmacology and Toxicology, KLE College of Pharmacy, KAHER, Belagavi, India for providing necessary facilities to conduct the work. The authors thank the Vamsi Labs Ltd. Solapur, India for providing Haloperidol and Micro Labs Ltd. Bangalore for providing L-DOPA as a gift sample.

Funding

This study has not having Funding from any Agencies.

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Authors and Affiliations

  1. Department of Pharmacology and Toxicology, KLE College of Pharmacy, KLE Academy of Higher Education and Research, Belagavi, Karnataka, 590010, India

    Aishwarya Vannur & Prakash R. Biradar

  2. ICMR-National Institute of Traditional Medicine, Belagavi, India

    Vishal Patil

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  1. Aishwarya Vannur
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  2. Prakash R. Biradar
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Contributions

Prakash R. Biradar supervised the experiments, confirmed the findings, helped in data analysis, helped in the manuscript writing. Aishwarya Vannur design the study, conducted the experiment, collected the data, drafted the manuscript. Vishal S. Patil statistical analysis and helped in manuscript drafting.

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Correspondence to Prakash R. Biradar.

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Ethical statement

The study protocol was reviewed and approved by the Institutional Animal Ethical Committee, KLE College of Pharmacy, KAHER, Belagavi, and Resolution No. KLECOP/CPCSEAReg. No. 221/Po/Re/S/2000/CPCSEA, Res. 25–13/10/2020. The animal experiments were carried out in accordance with the CPCSEA guidelines.

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Vannur, A., Biradar, P.R. & Patil, V. Experimental validation of Vitex negundo leaves hydroalcoholic extract for neuroprotection in haloperidol induced parkinson’s disease in rat. Metab Brain Dis 37, 411–426 (2022). https://doi.org/10.1007/s11011-021-00878-2

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