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  • The Internet Journal of Pharmacology
  • Volume 12
  • Number 1

Original Article

Hepatoprotective Effect Of ADHATODA VASICA Leaves Extract Against Paracetamol Induced Hepatic Damage In Rats

S Baishnab, V Satija, S Das

Keywords

adhatoda vasica, hepatoprotective and hepatotoxicity, paracetamol

Citation

S Baishnab, V Satija, S Das. Hepatoprotective Effect Of ADHATODA VASICA Leaves Extract Against Paracetamol Induced Hepatic Damage In Rats. The Internet Journal of Pharmacology. 2013 Volume 12 Number 1.

Abstract

The present study was conducted to evaluate the hepatoprotective activity of aqueous extract of Adhatoda vasica leaves against paracetamol induced hepatic damage. The aqueous extract of Adhatoda vasica leaves (100mg/kg & 200mg/kg) was administered orally for six days to the animals with hepatotoxicity induced by single dose of paracetamol (2gm/kg). Silymarin (25mg/kg) was given as reference standard. All the test drugs were administered orally. The aqueous extract of Adhatoda vasica leaves  in the doses of 100mg/kg, 200mg/kg body wt, reduced the levels of serum SGPT, SGOT and increased the Total protein and albumin/globulin ratio level significantly (p<0.01). Thus, the leaves of Adhatoda vasica have significant hepatoprotective activity.

 

Introduction

The liver plays a central role in transforming and clearing chemicals and is susceptible to the toxicity from these agents. Hepatotoxicity implies chemical-driven liver damage. Certain medicinal agents when taken in overdoses and sometimes even when introduced within therapeutic ranges may injure the organ. Other chemical agents such as those used in laboratories and industries, natural chemicals (e.g. microcystins) and herbal remedies can also induce hepatotoxicity. Chemicals that cause liver injury are called hepatotoxins. More than 900 drugs have been implicated in causing liver injury and it is the most common reason for a drug to be withdrawn from the market. Chemicals often cause subclinical injury to liver which manifests only as abnormal liver enzyme tests. Drug induced liver injury is responsible for 5% of all hospital admissions and 50% of all acute liver failures.1

The plant Adhatoda vasica Nees (AV) of the Acanthaceae family has been used for thousands of years in India. . Extracts of the leaves of AV are extensively used in cough, asthma, bronchitis, tuberculosis, inflammation and allergy.2-4  AV has been reported to posses, hepatoprotective activity. 5 The major bioactive chemical constituents of AV are the alkaloids like Vasicine, Vasicinone, Vasicinol, Vasicol, Vasicinolone, Vasicinine and adhatodine etc.6-7 The extract of A. vasica, also contain  high amount of flavonoid. 8  Keeping this in view, the present study was aimed at evaluating the hepatoprotective effect of Adhatoda vasica leaves Extract against Paracetamol Induced Hepatic Damage in Rats.

Materials and Methods

Plant material and extraction

The fresh leaves of AV were collected in the month of April to May from nagaon, Assam, India and duly authenticated. . The plant material was authenticated by Dr S.S.Dhawan, Professor of food and nutrition department, HAU, Hisar, Haryana.

The fresh leaves of AV were manually plucked, air dried, powdered (800 g) and aqueous extracts were prepared using sufficient water by percolation method followed by steam evaporation. A final yield of 20 g of the extract was obtained.9

Animals

Healthy adult Wistar albino rats (Rattus norvegicus) weighing 200—250 grams each were used for the study. All the animals were taken care of under ethical consideration as per the guidelines of the CPCSEA with due approval from the Institutional Animal Ethical Committee (Registration no: 634/02/a/CPCSEA; dated 19/5/2002).

Chemicals used

Crude powder of silymarin was obtained from MicroLabs India, Paracetamol was procured from Nulife Pharmaceuticals, India. Enzymes like alanine transaminase (ALT), aspartate transaminase (AST), total protein and albumin/globulin ratio were assayed using standard kits available. 

Experimental design for hepatoprotective study 10

A total of thirty animals were equally divided into five groups with six animals in each group:
Group–A: Normal Control - Received normal saline, 2 ml/kg/day                                                   

Group- B: Normal saline (2ml/kg) for 6 days + single dose of Paracetamol (2gm/kg) orally on  day 7

Group–C: Standard group treated with Silymain 25 mg/kg orally for 6 days + single dose of  Paracetamol (2gm/kg) orally on day 7                                                                                                                                                                                           Group–D: Test group treated with aqueous extract (200 mg/kg) orally for 6 days + single dose of Paracetamol (2gm/kg) orally on day 7.                                                                                                      

Group–E: Test groups treated with aqueous extract (100 mg/kg body weight) orally for 6  days + single dose of Paracetamol (2gm/kg) orally on day 7.

Induction of hepatotoxicity 10

Leaving aside six rats for Normal Control Group, 24 rats were induced hepatotoxicity by a single intraperitoneal injection of paracetamol in the dose of 2 gm/kg body weight. After 48 hours of treatment, blood was collected by intracardiac puncturing for biochemical markers.

Statistical analysis

The data was statistically analysed using One-way ANOVA.11 Values of p < 0.01 were considered significant.

Results

Paracetamol induced liver toxicity

The data was statistically analysed using One-way ANOVA. After 48 hr of administration of PCM, the serum levels of ALT, AST, were markedly increased whereas Plasma protein and Albumin/Globulin ratio were decreased when compared to normal control. Pretreatment with low dose of AV (100 mg/kg, p.o), medium dose of AV (200 mg/kg)  and silymarin significantly reduced the serum levels of ALT, AST, and increased  Plasma protein and Albumin/Globulin ratio when compared to PCM control (P<0.01)-(Table-1)

Table 1

Hepatoprotective effect of AV pretreatment on paracetamol induced in rats

Values are expressed as Mean ± SEM; n=6 rats in each group. One-way ANOVA done.  p<0.01 when compared to Normal Control Group.

Discussion

From the study, it was seen that pretreatment with low dose of AV (100 mg/kg, p.o), medium dose of AV (200 mg/kg) and silymarin significantly reduced the serum levels of ALT, AST, and increased Plasma protein and Albumin/Globulin ratio when compared to PCM control.

PCM is a commonly used as analgesic and antipyretic drug and is safe in therapeutic doses but produces fatal hepatic necrosis with toxic doses. 12 The toxic effect of PCM is due to oxidative damage induced by its metabolite, N-acetyl-p-benzoquinoneimine, produced by the action of cytochrome P-450 in the liver. This metabolite reacts with reduced glutathione (GSH) to yield non-toxic 3-GS-yl-PCM. Depletion of GSH causes the remaining quinone and other natural endogenous oxygen species to bind to cellular macromolecules leading to cell death.13  Damage induced by PCM is accompanied by an increase in the levels of serum biomarker enzymes.     

Flavonoids, tannins and microelements have been suggested to act as antioxidants and exert their antioxidant activity by scavenging the lipid peroxidation.14  The high scavenging property of Adhatoda vasica may be due to hydroxyl groups existing in the phenolic compound’s chemical structure that can provide the necessary component as a radical scavenger.15  Earlier studies conducted, revealed that vasicine content in Adhatoda vasica decreases significantly lipid peroxidation and similarly increases in antioxidants like superoxide dismutase, catalases, glutathione peroxidation and reduced glutathione.16

Conclusion

The present study reveals the hepatoprotective activity of AV against PCM induced hepatic damage in rats. The results show that AV is effective in low and medium doses (100 mg/kg, p.o and 200 mg/kg, p.o. Though the aqueous extract of Adhatoda vasica leaves showed significant hepatoprotective activity against Paracetamol induced hepatotoxicity, it is also needed further research to isolate the compound and exact mechanism responsible for hepatoprotective activity of the plant to rationalize its use as a drug to give more emphasis on this plant for the development of medicinal value.

References

1 Friedman, Scott E, Grendell, James H, McQuaid, Kenneth R, Current diagnosis & treatment in gastroenterology, Lang Medical Books/McGraw-Hill., New York, 2003; 664–679.
2 Dhuley JN. Antitussive effect of Adhatoda vasica extract on mechanical or chemical stimulation-induced coughing in animals. J Ethnopharmacol 1999;67:361-5.
3 Grange JM, Snell NJ. Activity of bromhexine and ambroxol, semi-synthetic derivatives of vasicine from the Indian shrub Adhatoda vasica, against Mycobacterium tuberculosis in vitro. J Ethnopharmacol 1996; 50:49-53.
4 Chakraborty A, Brantner AH. Study of alkaloids Adhatoda vasica Nees on their antiinflammatory activity. Phytother Res 2001; 15:532-4.
5 Bhattacharyya D, Pandit S, Jana U, Sen, S, Sur TK. Hepatoprotective activity of Adhatoda vasica aqueous leaf extract on D-galactosamine-induced liver damage in rats. Fitoterapia 2005 Mar; 76(2):223-25.
6 Bhatt VS. “Adhatoda vasica Nees – An Ayurvedic Medicinal Plant”. Indian Drugs 1978; 15(4):62-65.
7 Dr. Rajpal. Standardization of Botanicals. Eastern publishers New Delhi 2005; 2:1.
8. Sunita Maurya and Dhananjay Singh. Quantitative analysis of flavonoid in Adhatoda vasica Nees extracts. Der Pharma Chemica. 2010, 2 (5): 242-246
9. Pharmaceutical Formulas. Published at the Chemist and Druggist, London. 11th Edn. 1950; 1:183-185
10. Dhrubajyoti Sarkar et.al. Study the biochemical parameters and histopathological changes in liver of albino rats to find out the effect of methanolic extract of mimosa pudica leaves against paracetamol induced hepatic damages. The Pharma Research Journal, Year 2011, Volume 06, Issue 01, Page 50-57
11. Chiplonkar SA, Rao KV. Analysis of variance. In: Rao KV, editor. Biostatistics: a manual of statistical methods for use in health, nutrition and anthropology. 1st Edn. New Delhi: Jaypee Brothers, Medical Publishers (P) Ltd.; c1996. p. 237–72.
12. Mitchell JR, Jollow DJ, Potter WZ, Gillettee JR, Brodie BN. Acetaminophen-induced hepatic necrosis. I. Role of drug metabolism. J Pharmacol Exp Ther. 1973; 187: 185-88.
13. Udem SC, Madubunyy I, Okoye JOA, Anika SM. Anti-hepatotoxic effects of the ethanolic extracts of Combretum dolichopetalum root bark and Morinda lucida leaf. Fitoterapia. 1997; 68: 21-26
14. Yuting C, Rongliang Z, Zhongjian J, Yong J. Flavonoids as superoxide scavengers. Free Radical Biol Med 1990; 9:19-23.
15. M. Kessler, G. Ubeaud and L. Jung, J. Pharm and Pharmacol, 2003; 55, pp. 131–142
16. D.Srinivasarao et al. A study on antioxidant and anti-inflammatory activity of vasicine against lung damage in rats. Indian J Allergy Asthma Immunol 2006; 20(1):1-7.

Author Information

Seema Baishnab, MD, Asst Prof
Department of pharmacology, MAMC
Agroha, India
seemabaishnab@yahoo.co.in

Vikram Satija, MD, Associate Prof
Department of Paediatrics, MAMC
Agroha, India

Swarnamoni Das, MD, Professor and Head
Department of Pharmacology, Assam Medical College & Hospital
India

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