The Protective Effects Of Resveratrol On Liver Injury Induced By Isoniazid And Rifampicin In Rats And Its Mechanism

Objective: Resveratrol,a natural polyphenol compound mainly found in grapes,mulberry,polygonum cuspidatum and other plants,with antioxidant,anti-inflammatory,anti-cancer,anti-mutation and other biological functions.Many studies have demonstrated that resveratrol can prevent or slow down the progression of a wide variety of illness.It has been reported to have a protective effect on hepatic injury induced by ethanol,acetaminophen and et al.So far,there are few reports about its possible influence on liver injury induced by anti-TB drugs.Therefore,we intended this study to explore the hepatic protective effects of resveratrol on INH-RFP induced liver injury in rats and to investigate whether the protective mechanism is related to antioxidant,anti-inflammatory,and the expression of key genes and proteins in lipid metabolism.Methods: Sixty male Wistar rats,weighting 180~220g,which were randomly divided into five groups(n=12)including control group(Control),model group(Model),low-dose of resveratrol group(RLDG),middle-dose of resveratrol group(RMDG),high-dose of resveratrol group(RHDG).Low,middle,high dose resveratrol groups were respectively given resveratrol with doses of 25mg/kg,50mg/kg,100mg/kg dissolved in 0.5% CMC-Na.After two hours,excluding the rats in the control group,the other rats were given isoniazid 100mg/kg and rifampicin 150mg/kg by oral administration,control group was given equal dose of solvent intragastric perfusion.After continuous intragastric administration for 28 days,the rats were weighted and anesthetized with 3% pentobarbital sodium for 12 hours after the last treatment.The blood samples were collected by aorta ventralis puncture to determine biochemical parameters,and calculated the organ index.Histopathological examination were used to further demonstrate the changes of liver.Serum liver function and the level of lipid metabolism were measured by automatic biochemical analyzer.The activities of superoxide dismutase(SOD)in liver tissue was measured by using xanthine oxidase method;the concentration of malondialdehyde(MDA)was tested by using TBA;the glutathion(GSH)content and glutathione peroxidase(GSH-Px)activity were tested with DTNB,and the activities of catalase(CAT)in liver tissue was determined by visible spectrophotornetry method.The levels of TNF-?,IL-1?,IL-4 and IL-10 were detected with ELISA.The real-time PCR(q PCR)method was used to detect PPAR-?,PPAR-? m RNA expression levels and western blotting was used to determine protein expression levels of PPAR-?,PPAR-?.Results: 1.Compared with normal control group,the body weight of rats in model group and resveratrol group was well decreased and liver index was obviously increased.The levels of serum total bile acid(TBA),direct bilirubin(DBIL),total bilirubin(TBIL),alanine aminotransferase(ALT),aspartate aminotransferase(AST)and alkaline phosphatase(ALP)were increased significantly in the model group(P<0.01).Resveratrol groups significantly inhibited the elevation of TBA,DBIL,TBIL,AST which induced by isoniazid and rifampicin.High dose of resveratrol group could inhibit the increasing of serum activity of ALT.Liver histological analysis showed severe inflammatory cell infiltration and fatty degeneration in the model group,which were significantly improved in resveratrol groups.2.The level of lipid metabolism showed that the levels of serum CHOL,TG and liver CHOL were significantly increased(P<0.05),resveratrol intervention group successfully inhibited the elevation of CHOL,TG which induced by isoniazid and rifampicin.Compared with control group,the level of PPAR-?,PPAR-? m RNA and protein significantly up-regulated in liver of isoniazid-rifampicin treated rats(P<0.05).Resveratrol supplement group could obviously increase the m RNA and protein expression of PPAR-?,PPAR-?.3.INH-RFP produced a increase in MDA level as compared with normal control group,administration with resveratrol inhibited this increase.But no significant difference of MDA level existed among the groups.The hepatic GSH content and SOD activities were decreased in the model group.Low and high dose of resveratrol group could significantly increase the content of GSH in liver homogenate(P<0.05).The activity of GSH-Px and CAT in model group obviously went down.Middle dose of resveratrol group led to a prominent increase in GSH-Px and CAT activities,which were 20.6% and 53.23% higher than model group respectively(P<0.05).4.Serum TNF-? level in the model group were obviously higher than that in the control group,while the level of IL-10 was significantly decreased(P<0.05).Compared with model group,the level of serum TNF-? in the low,middle and high dose of resveratrol groups had decreased by 18.7%,15.5% and 28.6%(P<0.01).In addition,the level of IL-10 was both significantly increased in low and middle dose of resveratrol group.Conclusion: 1.Resveratrol has significant protective effects on INH-RFP induced liver injury.2.The underlying mechanisms of resveratrol on INH-RFP induced liver injury may be due to its ability of increasing the activity of antioxidant enzymes,inhibiting of inflammatory cytokines,regulating the level of lipid metabolism and up-regulation the expression of PPAR-?,PPAR-?.