JOURNAL CLUB

2023

Faculty of Pharmaceutical Sciences

PES University
JOURNAL CLUB
2023

Title :-Evaluation of the neuroprotective activity of P.

amarus in attenuating
arsenic-induced neurotoxicity - an in vivo study
Name of the Presenter :- Chaitra .R

Guide :- Dr. MUKUND HANDRAL

Faculty of Pharmaceutical Sciences

PES University
JOURNAL CLUB
2023

Title :-Evaluation of the neuroprotective activity of P.

amarus in attenuating
arsenic-induced neurotoxicity - an in vivo study
Authors :A. Hashima et.al
Article ID :: 100316
Publisher: Elsevier B.V
Journal: Phytomedicine

Faculty of Pharmaceutical Sciences

PES University
JOURNAL CLUB
Contents

1. Abstract
2. Introduction
3. Methodology
4. Results and discussion
5. Conclusion
6. References
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Abstract

Arsenic is frightful element in human history. Phyllanthus amarus is an

important medicinal herb proved to have neuroprotective effect in
experimental animals. This pre-clinical study used an animal model of
arsenic toxicity and screened the different neuroprotective mechanisms of
Phyllanthus amarus ethanolic extract.
Purpose: To assess the effect of Phyllanthus amarus ethanolic extract on
various biochemical parameter’s in the brain of arsenic administered
Wistar albino rats.
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Abstract

Methodology: The experimental animals (n=24) were divided into four

groups of 6 rats each. The animals received specific treatments .
At the end of the experiment rats were subjected to various
neurobehavioral tests such as Elevated plus maze, Light dark area, Forced
swim test and Tail suspension test.
After 24 h of the last treatment, the animals were euthanized and brain
tissue was assayed for various biochemical parameters
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Abstract

Results : The behavioral and biochemical data showed co-administration of

P. amarus ethanolic extract significantly reversed anxiety and depression-
like behaviors in experimental animals. EEPA effectively reduced acetyl
cholinesterase activity in the brain, and increased the levels of dopamine
and serotonin in the brains of treated animals. Arsenic treated significantly
decreased lipid peroxidase and increased brain catalase and SOD.
Conclusion: P. amarus have shown protective effects against arsenic
induced neurotoxicity.
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Introduction

Chronic arsenic poisoning due to contaminated subsoil water is a threat to

society. Arsenic contamination of drinking water from various sources has
been reported worldwide including both developed and developing
countries. Arsenic crosses the blood-brain barrier and has a wide range of
effects on brain white matter.
Brain damage is mainly due to oxidative stress , the treatment which are
available they do not have neuroprotective effect . Hence the plant with anti-
oxidants activity has increased with reducing the free radical and better
clinical recovery.
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Introduction

P. amarus is one of the important plants in Ayurveda used for the treatment of
diseases affecting the stomach, GI system, liver, kidney, and spleen. The
presence of a number of active compounds in P. amarus extract including
quercetin, a potential natural antioxidant that is an even more powerful
antioxidant than vitamin E.
The present study was carried out to assess the protective effect of Phyllanthus
amarus ethanolic extract in arsenic-induced CNS disorders by assessing various
biochemical parameters in Wistar albino rats.
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Methodology

1.Preparation of extract- 350g of the dry powder in soxhlet extractor apparatus

and was extracted with ethanol 90%.
2.Chemicals -Sodium arsenite
3.Experimental animals- Adult Wistar rats of either sex , weighing 150-170 g.
The animals were maintained according to the IAEC approved guidelines and
protocols in an air-conditioned animal house with constant 12 h light and 12 h
dark cycles. Animals were fed on standardized pellets for rodents and water ad
libitum.
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Methodology

5. Experimental protocol
GROUPING DRUGS VEHICLE DURATION
GROUP 1 CONTROL Distilled water ad 28 days
libitum
GROUP 2 Sodium arsenite salt 40 Drinking water 28 days
mg/kg b.w
GROUP 3 PAEE(100mg/kg po)+ Drinking water 28 days
sodium arsenite

GROUP 4 PAEE(200mg /kg po)+ Drinking water 28 days

sodium arsenite
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Methodology

On the 29th day, the rats were subjected to neurobehavioral

experiments such as EPM, LDA, FST and TST. The whole brain was
carefully removed and dissected out on a cold plate and it was placed in
phosphate buffer and refrigerated at – 20 ◦C and analyzed for
biochemical parameters.
Neurobehavioral experiments:
1. Elevated plus maze (EPM) 3. Forced swim test (FST)
2. Light and dark arena (LDA) 4. Tail suspension test (TST)
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Methodology

Biochemical estimations :
Estimation of brain
 Acetyl cholinesterase (AChE) enzyme determination
 Estimation of dopamine (DA)
 Estimation of serotonin (5-HT)
Estimation of antioxidants
 Superoxide dismutase (SOD)
 Catalase (CAT)
 Lipid peroxidase (LPO)
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Results and Disscuions

EPM test: PAEE decreased the time spent in closed arm

 Protective effect of PAEE on anxiety using EPM  Protective effect of PAEE on anxiety using LDA
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Results and Disscuions

 Protective effect of PAEE on depression using TST

 Protective effect of PAEE on depression using FST

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Results and Disscuions
Estimation of acetylcholinesterase level. Estimation of dopamine level

Estimation of serotonin level

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Results and Disscuions

Estimation of CAT level Estimation of SOD level

Estimation of LPO level

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Results and Disscuions

CONCLUSION:
The arsenic treated rats showed prominent anxiety and depression-like
behaviors in pharmacological tests. PAEE (100 mg/kg b.w and 200 mg/kg b.w)
significantly reversed anxiety and depression-like behaviors in experimental
animals.PAEE reduced AchE activity in the brain, increased the levels of DA and
5-HT of arsenic treated animals. A significant decrease in the activities of
antioxidant enzymes CAT, SOD, and increased the levels of LPO. Based on the
pharmacological, and biochemical analysis the plant extracts of P. amarus have
shown protective effects against arsenic induced toxicity.
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References

1. Ambali, S.F., Makinde, A.O., Shittu, M., Adeniyi, S.A., Mowuogwu, F.O., 2012. Alleviating
effect of Phyllanthus niruri on sensorimotor and cognitive changes induced by subacute
chlorpyrifos exposure in wistar rats. Am. J. Med. Med. Sci. 2, 50–58.
2. Amoateng, P., Quansah, E., Karikari, T.K., Asase, A., Safo, D.O. 2018. Medicinal Plants Used
in the Treatment of Mental and Neurological Disorders in Ghana. Evid Base Complement
Alternat Med. Available from: https://doi.org/10.1155/2018/8590381
3. Rahman, H., Eswaraiah, M.C. 2008. Simple Spectroscopic Methods for Estimating Brain
Neurotransmitters, Antioxidant Enzymes of Laboratory Animals Like Mice: A Review,
Available from: https://www.pharmatutor.org/articles/simple-spectroscop ic-method-
estimating-brain-neurotransmitter-antioxidnat-enzymes-lab-animals.
 THANK YOU
Department of pharmaceutical science
PES University