Hepatoprotective Activity of Cassia fistula Linn. Bark Extracts against Carbon Tetra Chloride Induced Liver Toxicity in Rats
G Parthasarathy, V Prasanth
Keywords
carbontetrachloride, cassia fistula, hepatoprotective
Citation
G Parthasarathy, V Prasanth. Hepatoprotective Activity of Cassia fistula Linn. Bark Extracts against Carbon Tetra Chloride Induced Liver Toxicity in Rats. The Internet Journal of Pharmacology. 2008 Volume 6 Number 2.
Abstract
In the present study cassia fistula (Caesalpinaceae) bark was extracted with water. The extracts were vacuum dried to yield Aqueous Extract (AQET). The extracts were evaluated for hepatoprotective activity against Carbon tetrachloride (CCl4) induced liver damage at 250 mg/kg and 500 mg/kg dose level. The biochemical parameters observed in serum were total bilirubin, Alkaline phosphatase (ALP), Serum glutamate oxaloacetate transaminase (SGOT), Serum glutamate pyruvate transaminase (SGPT) levels and total protein. Aspartate transaminase (AST), Alanine Transaminase (ALT) and total protein levels in liver were also evaluated. Histopathological study on the liver tissue was also performed. The extracts exhibited dose dependant reduction in total bilirubin, ALP, SGOT, SGPT, AST, ALT and increase in total protein (serum and liver) levels. The extract treated groups shows mild hepatatocytic damage compared to the CCl4 treated group. CONCLUSION:
Introduction
Cassia fistula linn (Caesalpinaceae) tree is one of the most widespread in the forests of India, usually occurring in deciduous forests The whole plant possesses medicinal properties useful in the treatment of skin diseases, inflammatory diseases, rheumatism, anorexia and jaundice 3 (Anonymous,1992 1 , Kirtikar and Basu 1991 2 ). A new bioactive flavone glycoside 5,3',4'- tri-hydroxy -6- methoxy -7-O- alpha -L- rhamnopyranosyl- (1-->2) -O- beta-D-galactopyranoside with antimicrobial activity was reported by (Yadava and Verma, 2003) 69 . Four new compounds, 5-(2-hydroxyphenoxymethyl)furfural, (2'S) -7- hydroxyl -5- hydroxymethyl -2- (2'-hydroxypropyl) chromone, benzyl 2-hydroxy-3,6-dimethoxybenzoate, and benzyl 2beta-O-D-glucopyranosyl-3,6-dimethoxybenzoate, together with four known compounds, 5-hydroxymethylfurfural, (2'S)-7- hydroxy-2-(2'-hydroxypropyl) -5- methylchromone, and two oxyanthraquinones, chrysophanol and chrysophanein, were also isolated from the seeds of Cassia fistula by Kuo et al. 5 , (2002). The heaptoprotective activity of leaf extracts (Bhakta et al., 1999; Bhakta et al., 2001) 3 and the hypoglycaemic activity have been reported. However, a detailed pharmacological screening of the Cassia fistula bark extracts have not been reported. The present study reports the Haepatoprotective activity of Cassia fistula bark extracts. In the present study C.
Materials And Method
Plant Material and Extraction
The barks of
Animals
Inbred wistar albino male rats (100 – 120gm) were used for the evaluation of pharmacological activities. They were kept in colony cages at 252 ° c, relative humidity 45 –55% under 12 h light and dark cycles. All the animals were acclimatized for a week before use. They were fed with standard animal feed (Hindustan Lever Ltd) and water adlibitum. The test compounds and the standard drugs were administered in the form of a suspension using 5% acacia as vehicle, to each group consisted of six animals.
All the pharmacological experimental protocols were performed according to the recommendation of the institutional animals ethics committee.
Acute oral toxicity
Acute oral toxicity test was performed as per OECD-423 guidelines (acute toxic class method). Wister albino mice (n=3) of either sex selected by random sampling technique were used for the study. The animals were kept fasting for 3-4 hours providing only water, after which the extracts were administered orally at the dose level of 5 mg/kg by intra gastric tube and observed for 3 days. If mortality was observed in 2-3 animals then the dose administered was assigned as toxic dose. If mortality was observed in one animal, then the same dose was repeated again to confirm the toxic dose. If mortality was not observed, the procedure was repeated for further higher doses such as 50, 300, and 2000 mg/kg.
Hepatoprotective activity
Wistar albino male rats were selected by random sampling technique. Rats were divided into four groups, each comprising of six rats. The groups were named as Group I (control), Group-II (CCl4 treated), Group-III (CCl4 +AQET 250 mg/kg treated) and Group-IV (ccl4 + AQET 500 mg/kg treated) and Group-IV (CCl4 + Silymarin 25 mg/kg treated). For the fist seven days of study Group-I and II were fed only with normal lab feed and water. Group-III and IV animals were treated orally with AQET 250 mg/kg/day and 500 mg/kg/day respectively for seven days and Group-V animals were treated with Silymarin (25 mg/kg/day).
On the seventh and eighth day animals of Group II, III, and IV & V were administered orally with a single dose of CCl4 with 5% acacia mixture (600 mg/kg/day). After thirty minutes of CCl4 administration Group III and Group IV rats were treated with AQET 250 and 500 mg/kg/day and Silymarin (25 mg/kg/day) respectively. All the animals were sacrificed by cervical decapitation under light ether anesthesia on the Ninth day. Blood was collected from jugular veins and centrifuged (300 rpm for 10 mins) to obtain serum. The serum was used for the assay of total bilirubin 17 , alkaline phosphatase (ALP) 16 , serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT) (Reitman and Frankel. 1957) 19 , and total protein 18 .
The Liver was dissected out immediately after sacrifice, washed in ice-cold saline. Small pieces of liver tissue were collected and preserved in 10% formalin solution for histopathological studies.
Statistical Analysis
The statistical analysis was carried out using one way analysis of variance (ANOVA) followed by Dunnet’s t-test.
Results and Discussion
Figure 1
The extracts did not cause mortality up to 2000 mg/kg and were considered as safe. ALP, SGOT, AST, ALT, and total bilirubin were significantly increased and total protein (serum and liver) was significantly decreased in CCl4 treated group. The extracts exhibited dose dependent reduction in total bilirubin, ALP, SGOT, SGPT, AST, ALT and increase in total protein (serum and liver) levels (Table 1).
Histology of liver from normal control group (Fig.1a) showed the central vein surrounded by cords of hepatocytes and showed normal arrangement of hepatocytes with clearly brought out nuclei, cytoplasm, central vein and portal triad. Microscopical examination of CCl4 treated liver (Fig 1b) showed cloudy swelling, sinusoidal dilatation, Individual hepatocytic necrosis of hepatic cells and centrilobular fatty changes with clear space representing fatty materials or lipids. AQET (250 mg/kg) treated animals (Fig. 1c) showed individual focal hepatocyte damage and necrosis. AQET (500 mg/kg) treated animals (Fig 1d) showed the mild focal hepatocytic damage and necrosis. The silymarin treated group (Fig 1e) showed central vein with cords of hepatocytes with occasional focal hepatocytic damage.
Figure 2
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Discussion
The reactive metabolites such as trichloromethyl (CCl3-) and trichloromethyl peroxy (CCl3OO-) radicals emanated from CCl4 initiate peroxidation of membrane unsaturated fatty acids. This lipid peroxidation of membrane seriously impairs its function and produces liver injury.
CCl4-induced damage produces alteration in the antioxidant status of the tissues, which is manifested as an abnormal histopathology like cloudy swelling, sinusoidal dilatation, Individual hepatocytic necrosis of hepatic cells and centrilobular fatty changes in Fig. (1b).
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