Hepatoprotective activity of aerial part of Glinus oppositifolius L. against Paracetamol-induced Hepatic Injury in Rats.

S. K. Sahu¹*, D. Das² and N. K. Tripathy¹

¹Department of Zoology, Berhampur University, Berhampur, Odisha –760007

²School of Pharmaceutical Sciences, SOA University, Bhubaneswar, Odisha-754030

*Corresponding Author E-mail: sujitkumar_2008@yahoo.com

ABSTRACT:

Ethanolic extract (80%) aerial part of Glinus oppositifolius was prepared and tested for its hepatoprotective effect against paracetamol-induced hepatitis in rats. Alteration in the levels of biochemical markers of hepatic damage like SGOT, SGPT, ALP, cholesterol and bilirubin were tested in both treated and untreated groups. Paracetamol (2 g/kg) has enhanced the SGOT, SGPT, ALP, cholesterol and bilirubin levels. Treatment with ethanolic extract of aerial part of G. oppositifolius (200 mg/kg and 400 mg/kg) has reverse back the altered levels of biochemical markers to the near normal levels in the dose dependent manner.

KEYWORDS: Glinus oppositifolius; Hepatoprotective activity; Paracetamol; Silymarin.

INTRODUCTION:

Glinus oppositifolius belongs to family Molluginaceae ¹, is an annual or perennial sub shrubs, or shrubs, rarely dioecious, glabrous or rarely hairy; Stems erect or prostrate; Stem simple, alternate, rarely opposite; Flowers bisexual, Petals absent or few to many, white, pink, or purple. Fruit usually a loculicidal capsule rarely breaking into 2 nutlets; Seeds with embryo curved around a hard, starchy perisperm ². Traditionaly G. oppositifolius is used in the treatment of skin disease, increase appetite, cures vata, kapha, piles, leucoderma, tonic to intestine, urinary infections, fever, cough, liver problem and also used as antioxidant due to its excellent properties and potent phytoconstituents ³. Activities like Free radical scavenging and Antioxidant activities of the ethanol extract ⁴. Hepatoprotective effect of a methanolic extract of root ⁵. Antiprotozoal activity of aerial part ⁶. Immunomodulating activity of aerial part of G. oppositifolius ⁷.

It has been reported, an amino acid derivative, L-(−)-(N-trans-cinnamoyl)-arginine, was isolated from the whole plant of G. oppositifolius (L.) Aug. DC. along with kaempferol 3-O galactopyranoside, is orhamnetin3-O-â-Dxylopyranosyl-(1→2)-â-D-galactopyranoside, vitexin, vicenin-2, adenosine and L-phenylalanine was reported⁸. The liver is a vital organ involved in the maintenance of metabolic functions and detoxification from the exogenous and endogenous challenges, like xenobiotics, drugs, viral infections and chronic alcoholism. Therefore, this present study was under taken to evaluate the hepatoprotective effect of the ethanolic extract of aerial part of the plant G. oppositifolius against paracetamol induced hepatitis in rats.

MATERIALS AND METHODS:

Plant material:

The plant G. oppositifolius was collected from the rural belt of Rayagada (Odisha) during the month of February and was authenticated by the taxonomist of Botanical Survey of India, Howrah. The collected plant materials were washed under running tap to remove adhered dirt, and then shed dried. Then the aerial part was ground in to coarse powder.

Preparation of extract:

The powered plant material was defatted with petroleum ether (60-80°c) and then extracted with 80% ethanol using soxhlet apparatus. The solvent was removed under reduced pressure to obtain dry extract, which gave a dark greenish-black coloured sticky residue. The extract was stored in desiccator for further use.

Phytochemical Screening:

In this research work the ethanolic extract of both G. oppositifolius was qualitatively tested for the presence of chemical constituents. It shows the presence of carbohydrates, alkaloids, gums, saponins, flavonoids and steroids^8,9.

Animals:

Swiss albino mice (20-25 g) of either sex were used for acute toxicity study and adult wistar albino rats (150-200 g) of either sex were used for evaluation of pharmacological studies. The animals were kept in standard polypropylene cages at room temperature of 34 ± 2 ͦC and at 60-65 % relative humidity during the experimental work. The experiment has been performed in the CPCSEA approved laboratory of Institute of Pharmacy and Technology, Salipur (Regd. No. 1053/ac/07/CPCSEA) with the permission of Institutional animal ethics committee.

Acute toxicity study:

The acute toxicity of ethanolic extract of G. oppositifolius was determined as per the CPCSEA guideline no. 420 (fixed dose method). It was observed that the test extracts shows no mortality even at 2000 mg/kg dose hence, 1/10^th (200 mg/kg) and 1/5th (400 mg/kg) of this dose were selected for further study.

Evaluation of hepatoprotective activity:

The method of Setty et al., 2007 was used in this study⁹. The wistar albino rats (150-200 g) of either sex were divided into five groups of six animals each. Group I received saline 1 ml/kg for one week (control). Group II received saline 1 ml/kg for one week (positive control). Groups III, received Silymarin (100 mg/kg p.o.). Group IV and V received 200 and 400 mg/kg of ethanolic extract of G. oppositifolius respectively, once a day for seven days. On the fifth day, after the administration of the respective treatments, all the animals of groups II, III, IV and V were administered with Paracetamol 2 g/kg orally. On the seventh day after 2 hours of respective treatments the blood samples were collected after scarification of animals by cervical dislocation for the estimation of biochemical marker enzymes¹⁰.

Biochemical studies:

The blood was obtained from all animals by cardiac puncture. The blood samples were allowed to clot for 45 min. at room temperature. Serum was separated by centrifugation 2500 rpm at 30ºC for 15 min. and utilized for the estimation of various biochemical parameters like SGOT and SGPT were measured according to the method of Rietman and Frankel¹¹. ALP was measured according to the method of King¹². Serum bilirubin and cholesterol were estimated following Malloy and Evelyn method¹³. The biochemical tests were done by using following methods with help of the kit supplied by Span Diagnostic Limited, G.I.D.C, Sachin-394230 (Surat), India.

Statistical analysis:

All the results were statistically analyzed using one way ANOVA followed by Dunnet's t-test. Values are expressed as mean ± S.E.M, (n=6). *P<0.05 and **P<0.01 compared with control was considered as significant.

Table No.1: Effect of the ethanolic extract of G. oppositifolius (EEGO) on biochemical parameters in Paracetamol (PCM) induced hepatic injury in rats.

Group	Treatment	Dose (mg/Kg)	SGOT(U/L)	SGPT(U/L)	ALP(U/L)	Cholesterol (mg/dL)	Total Bilirubin (mg/dL)	Direct Bilirubin (mg/dL)
I	Control	5 ml/Kg	132.26± 2.61	63.27± 2.91	139.76± 1.27	101.92± 1.35	0.90± 0.02	0.25± 0.01
II	PCM	2000	391.78± 3.93	277.80± 2.74	435.99± 2.16	162.51± 2.06	3.40± 0.06	0.68± 0.01
III	PCM+Silymarin	2000+ 100	138.26± 2.08**	81.45± 1.97**	159.62± 2.02**	116.50± 1.57**	1.03± 0.02**	0.26± 0.01**
IV	PCM+EEGO	2000+ 200	273.22± 4.64**	179.55± 3.42**	285.06± 1.42**	137.04± 2.39**	1.45± 0.01**	0.40± 0.01**
V	PCM+EEGO	2000+ 400	161.61± 3.70**	90.92± 2.04**	183.45± 2.13**	124.36± 1.58**	1.07± 0.01**	0.31± 0.01**

Values are expressed as mean ± S.E.M. (n=6). Significant at *P<0.05 and **P<0.01 compared with vehicle control (ANOVA followed by Dunnet’s t-test).

RESULT AND DISCUSSION:

Paracetamol (Acetaminophen) is a widely used antipyretic and analgesic, produces acute liver damage if overdoses are consumed. Paracetamol is mainly metabolized in liver to excretable glucuronide and sulphate conjugates¹⁴. However, the hepatotoxicity of Paracetamol has been attributed to the formation of toxic metabolites. Paracetamol produces hepatic necrosis when ingested in very large doses. The hepatoprotective activity of the ethanolic extract of G. oppositifolius was monitored by estimating serum enzyme levels of SGOT, SGPT, ALP, cholesterol and bilirubin which give a good idea about the functional state of the liver. As shown in table no.1 enzyme levels of SGOT, SGPT, ALP, cholesterol and bilirubin were markedly enhanced in Paracetamol treated animals compared to normal rats indicating liver damage. Administration of ethanolic extract of G. oppositifolius at a dose of 400 mg/kg, p.o., remarkably prevented Paracetamol induced increase of serum enzymes, cholesterol and bilirubin level. Treatment with the standard drug also reversed the hepatotoxicity significantly.

ACKNOWLEDGEMENTS:

The authors are very grateful to the Zoology department of Berhampur University, Berhampur and Institute of Pharmacy and Technology, Salipur, Cuttack, Odisha, India for providing required facilities of this work.

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Received on 09.11.2012 Accepted on 22.11.2012

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