艾迪注射液对DEN诱导型肝癌大鼠体内细胞色素P450酶表达的影响

Effect of Aidi Injection on Expression of Cytochrome P450 Enzyme in DEN-induced Hepatocellular Carcinoma Rats

目的:观察艾迪注射液(AD)对二乙基亚硝胺(DEN)化学诱导的原发性肝癌(HCC)大鼠体内细胞色素P450(CYP450)4种亚型酶CYP1A2,CYP2E1,CYP3A2,CYP2C11 mRNA和蛋白表达的影响。方法:健康SD雄大鼠随机选取3只作为空白组,余下大鼠采用DEN间断性诱导原发性肝癌大鼠模型,模型成功后随机将大鼠分为模型组,AD组,每组3只。正常组与模型组腹腔注射10 mL·kg^(-1)生理盐水,AD组腹腔注射10 mL·kg^(-1)AD,1次/d,共干预14 d。采用实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)分别检测CYP1A2,CYP2E1,CYP3A2,CYP2C11 mRNA和蛋白的表达。结果:Real-time PCR结果表明,给药14 d,与正常组比较,模型组大鼠癌旁组织(PCT)和癌灶组织(CT)CYP1A2,CYP2E1,CYP3A2,CYP2C11 mRNA表达均明显下调(P<0.05,P<0.01);与模型组比较,AD组PCT中4种亚型酶mRNA表达明显下调(P<0.05,P<0.01),CT中4种亚型酶mRNA表达明显上调(P<0.05)。Western blot结果表明,与正常组比较,模型组CT中CYP1A2,CYP2E1,CYP3A2,CYP2C11蛋白表达均显著下调(P<0.01),PCT中CYP3A2,CYP2C11蛋白表达显著下调(P<0.01);与模型组比较,AD组CT,PCT中CYP1A2,CYP2E1,CYP3A2,CYP2C11蛋白表达有下调趋势,差异无统计学意义。结论:AD可下调大鼠肝组织中CYP1A2,CYP2E1,CYP3A2,CYP2C11 mRNA和蛋白的表达。临床使用AD时,应注意可能因CYP450酶抑制引起的药物相互作用。

Objective: To study the effect of Aidi injection(AD)on the expression of cytochrome P450 isoenzyme 1A2,2E1,3A2,2C11(CYP1A2,2E1,3A2,2C11)mRNA and protein in rats with Nnitrosodiethylamine(DEN) chemically induced primary hepatocellular carcinoma(HCC). Method: Three healthy SD male rats were randomly selected as the blank group,and the remaining rats were treated with DEN intermittently induced primary hepatocellular carcinoma rat model. After success of the model,the rats were randomly divided into model group and AD group,with 3 rats in each group. The rats in the blank group and model group were intraperitoneally injected with 10 mL·kg^(-1) saline, while those in the AD group were intraperitoneally injected with 10 mL·kg^(-1) AD once a day,a total of 14 d intervention. Real-time quantitative polymerase chain reaction(Real-time PCR) and Western blot were used to detect the mRNA and protein expressions of CYP1A2,CYP2E1,CYP3A2 and CYP2C11,respectively. Result: Real-time PCR results showed that after 14 days of drug administration,compared with the blank group,the mRNA expressions of CYP1A2,CYP2E1,CYP3A2 and CYP2C11 were all down-regulated in para-cancerous tissue(PCT)and cancerous tissue(CT)in model group,and there were significant differences(P<0.05,P<0.01). Compared with the model group,the mRNA expressions of the four subtype enzyme were significantly down-regulated in PCT in the AD group(P<0.05,P<0.01),while the mRNA expressions of the four subtype enzyme were significantly up-regulated in CT(P<0.05),and the expression was down-regulated overall. Western blot results showed that compared with the blank group,the protein expressions of CYP1A2,CYP2E1,CYP3A2 and CYP2C11 in CT of the model group were significantly down-regulated(P<0.01),and the protein expressions of CYP3A2 and CYP2C11 were significantly down-regulated in PCT(P<0.01). Compared with the model group,the protein expressions of CYP1A2,CYP2E1,CYP3A2 and CYP2C11 in CT and PCT were down-regulated in the AD group,but the differences were not statistically significant. Conclusion:AD can down-regulate the mRNA and protein expressions of CYP1A2,CYP2E1,CYP3A2 and CYP2C11 in rat liver tissues. In clinical use of AD,attention should be paid to drug interactions that may be caused by CYP450 enzyme inhibition.