细胞色素P450 2E1在利福平减弱异烟肼引起肝脏发生氧化应激中的作用

The role of cytochrome P450 2E1 on rifampicin attenuating the isoniaizid-induced oxidative stress in liver

目的研究利福平(RIF)减弱异烟肼(INH)引起小鼠肝脏发生氧化应激的作用机制。方法将♀ICR成年健康小鼠随机分为4组:50mg/kgINH组;100mg/kgRIF组;INH+RIF组:同时给予INH(50mg/kg)和RIF(100mg/kg);对照组:给予等容量的溶剂,所有小鼠每日均给予处理1次,连续处理1周。分析血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(AKP)、总胆红素(TB)和总胆汁酸(TBA)水平,Griffith法测定肝脏组织谷胱甘肽(GSH)含量,TBARS比色法分析肝脏组织脂质过氧化水平,RT-PCR技术检测肝脏组织细胞色素P4502E1(CYP2E1)mRNA水平,Westernblot检测肝脏组织CYP2E1蛋白表达水平。结果 INH、RIF及RIF与INH共处理组小鼠血清中ALT均轻度升高,INH处理显著升高肝脏组织脂质过氧化水平并降低GSH含量,RIF共处理显著减弱INH升高肝脏组织脂质过氧化水平并降低GSH含量。单纯RIF处理显著下调肝脏组织CYP2E1蛋白表达水平,且RIF共处理显著减弱INH对肝脏组织CYP2E1蛋白表达的诱导作用;所有处理对CYP2E1mRNA水平无明显影响。结论 RIF减弱INH引起肝脏组织发生氧化应激中,CYP2E1发挥了一定作用。

Objective To investigate the mechanism of rifampicin attenuate the isoniaizid-induced oxidative stress in mouse liver. Methods Animals received anti-TB drugs,rifampicin and isoniaizid-alone or in combination-once daily for up to one week. Assays for alanine aminotransferase（ALT） ,alkaline phosphatase（ALP） were performed to assess liver toxicity. The GSH content in liver was determined by the method of Griffith and lipid peroxidation was quantified by measuring malondialdehyde（MDA） . The expressions of CYP 2E1 in liver were determined by Western blotting. Results Compared with the animals received INH alone,the group received rifampicin and isoniaizid in combination produced a progressive enhancement in glutathione （GSH） and a descendent in malondialdehyde （MDA） ,and there was no significant difference between the group received RIF alone and the group for contrast. RIF significantly reduced the expression of CYP 2E1 in mouse liver. Conclusion RIF can attenuate the INH induced oxidative hepatic injury via reduce the expression of CYP 2E1 and cause imbalance in endogenous enzymatic oxidant-antioxidant defense.