Protective Effects Of 70 Wei Zhen-Zhu-Wan On CCl₄ Liver Injury In Mice And Its Effects On Cytochrome P450

Objective:This study examined its protective effects against carbon tetrachloride?CCl₄?-induced liver injury in mice and possible mechanisms.Methods:?1?Mice were randomly divided into 5 groups,respectively as normal control,model control,70W(150,500 mg·kg^-1,p.o.×7 d),and oleanolic acid(25 mg·kg^-1,i.p.×3 d)as positive control.Twenty-four hr after the last dose,mice were intraperitoneally given 0.1%CCl₄(10 mL·kg^-1,18 h),and liver injury was examined.Toxicity-related gene expression was determined by RT-PCR.?2?Normal mice were given 70W(500mg·kg^-1,p.o.×4 d),the expression of antioxidation gene metallothionein 1?MT-1?,the Nrf2 antioxidant pathway genes Nrf2,Nqo1,Ho-1,Gclc,GST-mu and GST-pi was examined.?3?Mice were given 70W(0.15,0.5,1.5 g·kg^-1,p.o.),Zuotai(30 mg·kg^-1,p.o.),HgCl₂(33.6 mg·kg^-1,p.o.)and MeHgCl(3.1 mg·kg^-1,p.o.)for 7 days.Liver CYP450 mRNA and proteins were isolated for real-time RT-PCR,Western blot analysis.Results:?1?CCl₄ produced liver injury,as evidenced by increased serum enzyme activities and histopathology.Compared with model group,the high dose of 70W and oleanolic acid reduced the serum ALT and AST elevated by CCl₄ and improved liver histopathology.In 70W and oleanolic acid groups,liver cell degeneration,inflammation,and necrosis were reduced.The CCl₄ induced expression of DNA damage genes?Gadd45,Gadd153?and inflammation genes?MIP2 and IL-6?was reduced by 70W and oleanolic acid.?2?70W pretreatment induced the expression of liver detoxification genes such as MT-1 and the Nrf2 antioxidant pathway genes Nrf2,Nqo1,Ho-1,Gclc,GST-mu and GST-pi.?3?70W and Zuotai had no apparent effects on mRNA expression of hepatic AhR and Cyp1a2;CAR,Cyp2b10 and Cyp2e1;PXR and Cyp3a11;PPAR?and Cyp4a10;as well as FXR and Cyp7a1,while HgCl₂ and MeHgCl increased the expression of Cyp1a2,Cyp2b10,Cyp7a1.Western-blot analysis confirmed gene expression changes.Conclusion:70W protected mice from acute liver injury induced by CCl₄.The protection appears to be mediated via induction of metallothionein and Nrf2 antioxidant genes,rather inhibition of CCl₄ bioactivation via CYP2E1 and CYP1A2.70W had no apparent effect on the hepatic cytochrome P450 system under clinical doses.Its effects on P450 were different from HgCl₂ and MeHgCl.