目的:观察四氯化碳(carbon tetrachloride,CCl4)诱导的肝纤维化大鼠肝脏P450亚型CYP3A1表达的变化。方法雄性SD大鼠采用10%四氯化碳诱导肝纤维化模型(1ml/kg,2次/w,共12w),收集大鼠外周血和肝脏;石蜡切片的HE染色观察大鼠肝脏的病理学改...目的:观察四氯化碳(carbon tetrachloride,CCl4)诱导的肝纤维化大鼠肝脏P450亚型CYP3A1表达的变化。方法雄性SD大鼠采用10%四氯化碳诱导肝纤维化模型(1ml/kg,2次/w,共12w),收集大鼠外周血和肝脏;石蜡切片的HE染色观察大鼠肝脏的病理学改变;外周血离心获得血清后检测肝功能指标 ALT、AST;Real time RT-PCR检测肝纤维化大鼠肝脏组织中CYP3A1的mRNA表达。结果四氯化碳诱导的肝纤维化大鼠肝脏 HE染色可见明显纤维化改变,外周血的肝功能生化检测显示肝纤维化大鼠的ALT和AST较空白对照组显著升高(P<0.05);而Real time RT-PCR检测结果显示四氯化碳诱导的肝纤维化大鼠肝脏中CYP3A1 mRNA的表达较空白对照组明显降低(P<0.05)。结论肝药酶CYP3A1在四氯化碳诱导的肝纤维化大鼠肝脏中的表达是降低的。展开更多
Clinical drug-drug interactions（DDIs） induced by CYP3A may reduce the exposure and pharmacological activity of CYP3A substrate.Up-regulation of CYP3A mRNA is often used to evaluate inductive effect of test compounds...Clinical drug-drug interactions（DDIs） induced by CYP3A may reduce the exposure and pharmacological activity of CYP3A substrate.Up-regulation of CYP3A mRNA is often used to evaluate inductive effect of test compounds on CYP3A. A quantitative real time PCR assay was developed and validated for the absolute quantification of CYP3A1 and CYP3A2 mRNA.Specific primers of CYP3A1,CYP3A2 and GAPDH（glyceraldehyde-3-phosphate dehydrogenase,as a house-keeping gene） were well designed.The relationship between threshold cycle（Ct） and logarithm of the concentrations of CYP3A1, CYP3A2 and GAPDH was linear ranged from 1 attomol/μL to 1×10~6 attomol/uL with great inter- and intra-assay reproducibility. This method was successfully applied to investigate the time courses of CYP3A1 and CYP3A2 mRNA induction in rat liver after 100 mg/kg dexamethasone（DEX） administration by intraperitoneal（i.p.） injection.The baseline levels of CYP3A1 and CYP3A2 mRNAs were 37.78 attomol/ug（total RNA） and 252.31 attomol/ug（total RNA）,respectively.CYP3A1 and CYP3A2 mRNA values increased gradually to their peak levels（19- and 8- fold vs.baseline） within 24 h and 42 h,respectively,and then returned to their baseline 60 h after DEX administration.展开更多
文摘目的:观察四氯化碳(carbon tetrachloride,CCl4)诱导的肝纤维化大鼠肝脏P450亚型CYP3A1表达的变化。方法雄性SD大鼠采用10%四氯化碳诱导肝纤维化模型(1ml/kg,2次/w,共12w),收集大鼠外周血和肝脏;石蜡切片的HE染色观察大鼠肝脏的病理学改变;外周血离心获得血清后检测肝功能指标 ALT、AST;Real time RT-PCR检测肝纤维化大鼠肝脏组织中CYP3A1的mRNA表达。结果四氯化碳诱导的肝纤维化大鼠肝脏 HE染色可见明显纤维化改变,外周血的肝功能生化检测显示肝纤维化大鼠的ALT和AST较空白对照组显著升高(P<0.05);而Real time RT-PCR检测结果显示四氯化碳诱导的肝纤维化大鼠肝脏中CYP3A1 mRNA的表达较空白对照组明显降低(P<0.05)。结论肝药酶CYP3A1在四氯化碳诱导的肝纤维化大鼠肝脏中的表达是降低的。
基金National Integrity Innovational Technology Platform of New Drug and Development(Grant No.2009ZX09301- 010)
文摘Clinical drug-drug interactions（DDIs） induced by CYP3A may reduce the exposure and pharmacological activity of CYP3A substrate.Up-regulation of CYP3A mRNA is often used to evaluate inductive effect of test compounds on CYP3A. A quantitative real time PCR assay was developed and validated for the absolute quantification of CYP3A1 and CYP3A2 mRNA.Specific primers of CYP3A1,CYP3A2 and GAPDH（glyceraldehyde-3-phosphate dehydrogenase,as a house-keeping gene） were well designed.The relationship between threshold cycle（Ct） and logarithm of the concentrations of CYP3A1, CYP3A2 and GAPDH was linear ranged from 1 attomol/μL to 1×10~6 attomol/uL with great inter- and intra-assay reproducibility. This method was successfully applied to investigate the time courses of CYP3A1 and CYP3A2 mRNA induction in rat liver after 100 mg/kg dexamethasone（DEX） administration by intraperitoneal（i.p.） injection.The baseline levels of CYP3A1 and CYP3A2 mRNAs were 37.78 attomol/ug（total RNA） and 252.31 attomol/ug（total RNA）,respectively.CYP3A1 and CYP3A2 mRNA values increased gradually to their peak levels（19- and 8- fold vs.baseline） within 24 h and 42 h,respectively,and then returned to their baseline 60 h after DEX administration.
