Real-time PCR方法检测高脂饲喂小鼠肝脏CYP2A5的表达

Real-time fluorescence quantitative PCR assay for detection of high-fat feeding murine liver tissue CYP2A5 expression

为了建立非酒精性脂肪肝(NAFLD)模型小鼠肝组织中CYP2A5表达的实时荧光定量PCR(real-time fluorescence quantitative PCR,qRT-PCR)方法。高脂饮食诱导小鼠至8周,采用qRT-PCR方法检测小鼠肝脏中CYP2A5的表达水平,同时评价该方法的特异性。建立了小鼠肝组织中CYP2A5的SYBR Green实时荧光定量PCR检测方法,结果显示,该方法的溶解曲线为单峰,同时核酸电泳显示一条特异性条带。检测结果表明,高脂诱导的NAFLD小鼠肝组织中CYP2A5的表达水平极明显高于正常对照组(P<0.01)。表明该实时荧光定量PCR方法,特异性好、灵敏度强,为进一步研究小鼠CYP2A5的表达提供了试验基础。

In order to establish real-time fluorescence quantitative PCR method for detection liver CYP2A5 expression of nonalcoholic fatty liver disease （NAFLD）mouse model of NAFLD was induced with high fat diet for eight weeks. The relative expression level of mouse CYP2A5 in liver tissue was detected by qRT-PCR. The sensitivity and specificity of this method were evaluatd. The real-time fluorescence quantitative PCR method for detection liver CYP2A5 expression of NAFLD was established. The melting curve showed single peak and the amplified products were electrophoresed and a single product was observed. Compared to the control mice, the CYP2A5 relative expression was significantly increased in NAFLD mice liver tissue （P〈0.01）. Thus a standard method of qRT-PCR can be used for the detection of mouse CYP2A5 expression due to its good specificity and sensitivity.