Hepatoprotective and antioxidant effect of Sphaeranthus indicus against acetaminophen–induced hepatotoxicity in rats.
B Tiwari, R Khosa
acetaminophen, antioxidant activities, hepatocellular damage, hepatoprotection, sphaeranthus indicus
B Tiwari, R Khosa. Hepatoprotective and antioxidant effect of Sphaeranthus indicus against acetaminophen–induced hepatotoxicity in rats.. The Internet Journal of Tropical Medicine. 2009 Volume 6 Number 2.
The flower heads of
The pharmaceutical imbalance between remedies that protect the liver and have antioxidant properties and drugs that induce hepatotoxicity has prompted and accelerated research into plants used in folk medicines to treat liver diseases and boost liver functions.
Materials And Methods
Plant materials: Dried flower heads of
Extraction: Dried powdered flower heads were powdered mechanically (Sieve No. 10/44) about 250g of the powder was thougherly extracted with methanol in a soxhlet apparatus. Another 250g of powdered material was completely extracted in boiling distilled water. A rotary vacuum evaporator concentrated both the extracts the percentage yield of both the extracts 22.5% and 20.9% respectively. The residue of both extracts made into a suspension in water and propylene glycol (4:1) containing Tween-80 (0.08%) at the concentration of 200mg/ml separately.
Animals: Thirty pathogen free male albino rats (four weak) of either sex weighing 180-200g respectively were used for the study. They were housed in specific standard laboratory conditions and were kept in temperature control environment (25±10C), in a relative humidity (55± 5%), with regular 12h light/12h dark cycle. All animals were fed with standard rat chow diet, water
Hepatoprotective activity: Rats were divided into five groups, with six animals in each group. Group I, the normal control group animals were administered p.o., a single daily dose of 0.5% Tween-80 (1ml) on all five days. Group II, the APAP control animals were administered a single daily dose of 0.5%Tween 80 (1ml) p.o., on all the 5 days and on second and third day they were administered APAP (2g/kg p.o.,). Group III and IV animals were administered AQS and MES suspensions respectively (300mg/kg p.o.,) on all five days and a single dose of APAP (2g/kg p.o.,) on days second and third, 30 min after of each extracts. Group V animals were administered Silymarin, the known hepatoprotective compound (Sigma Chemical Company USA), at a dose of 100mg/kg p.o., on all 5 days and single dose of APAP (2g/kg p.o.,) on days 2 and 3,30 min after silymarin administration. The blood was withdrawn through retro-orbital plexus of rats on 5th day. Microscopic observation of liver was also done.
Assessment of antihepatotoxic activity:
Assessment of antihepatotoxic activity was done by determining the glutamyl pyruvate transaminase (SGPT) and glutamyl oxalacetic acid transaminase (SGOT) enzyme activity. The enzyme assay was carried out by Reagent Kits maintained by Biocon India Ltd Bangalore and the procedures were essentially those described in the literature available with kits. Estimations were made on Auto-analyser; Reitman and Frankel method (1957) was used for determining the enzyme activity in the supernatant of various groups.
On fifth day, animals were sacrificed and blood was collected directly through retro-orbital plexus. serum was separated after coagulating at 37 ◦C for 30 min and centrifuging at 1200–1500 rpm for 15–20 min. Serum was analyzed for various biochemical parameters, i.e. serum glutamyl oxalacetic acid transaminase (SGOT), serum glutamyl pyruvate transaminase (SGPT).12 acid phosphatase (ACP) and alkaline phosphatase (ALP).13 serum bilirubin (SB).14 and total protein.15
Determination of antioxidant enzyme activity: The liver was perfused with 0.86% cold saline to remove all the red blood cells. Then it was suspended in 10% (w/v) ice cold 0.1M phosphate buffer (pH 7.4) cut into small pieces, and required quantity was weighed and homogenized using a Teflon homogenizer. The homogenate was used for estimation of enzymic and non-enzymic antioxidants like SOD, 16 CAT, 17 GPx, 18 and lipid peroxide level.19
Results: Serum activities of transaminases, SGPT, SGOT and ACP, ALP, serum bilirubin and total protein are given in Table.1 where the single dose of APAP significantly elevated SGPT, SGOT activities when compared to normal animals. (Table.1).
Treatment of AQS & MES (300mg/kg) extracts 1h prior to APAP administration significantly protected the elevation of marker enzymes, serum bilirubin and ACP and ALP activities. Reduced activities of enzymic and non-enzymic antioxidants and enhanced activities of lipid peroxidation were seen in the APAP-treated group (Table.2), whereas standard silymarin and the drug treated group showed significant (p<0.01) rise in antioxidant level with reduction in lipid peroxidation level when compared with the standard drug, silymarin (Table. 2).
Effect of Sphaeranthus indicus flower heads on liver antioxidant enzymes and lipid peroxidation paracetamol intoxicated rats
Histopathology of Group II animals shows patches of liver cell necrosis with inflammatory collections around central vein where as drug treated group showed absence of cell necrosis but with minimal inflammatory conditions around the central vein. The MES (300mg/kg, p.o)-treated group showed minimal inflammatory conditions with near normal architecture possessing higher hepatoprotective action (Fig.3)
Discussion: In the present study the methanolic extract of
In conclusion the present study has demonstrated that methanolic extract of