大鼠无肝状态小肠CYP3A1酶活性增强和mRNA表达增高及芬太尼肝外代谢  

Activity and mRNA expression of cytochrome P450 and fentanyl metabolism in rat intestine in anhepatic phase

作　　者：李玉萍[1] 顾健腾[1] 陶国才[1] 向学凤[2] 杨波[2] 

机构地区：[1]第三军医大学西南医院麻醉科,重庆400038 [2]第三军医大学西南医院药理基地,重庆400038

出　　处：《第三军医大学学报》2008年第7期606-609,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30600585)~~

摘　　要：目的研究无肝期前后芬太尼血药浓度变化及其代谢酶CYP3A1在大鼠小肠活性的改变和基因表达的变化,探讨无肝期芬太尼的代谢及其形成原因。方法分两部分实验:实验一,20只大鼠随机分成对照组和阻断肝门组(n=10),LC/MS/MS法测芬太尼血药浓度,观察无肝期前后芬太尼的代谢;实验二,30只大鼠随机分为3组(n=10):①为对照组(B1组);②阻断肝门30min组(B2组);③阻断肝门60min组(B3组)。采用逆转录聚合酶链式反应(RT-PCR)检测各组CYP3A1 mRNA的表达,荧光比色法检测CYP3A1酶的活性。结果无肝期组芬太尼浓度下降比对照组明显减慢(P<0.05),但仍有下降趋势。小肠组织CYP3A1酶的活性和mRNA表达水平B2、B3组均明显高于B1组。B2组与B3组差异无统计学意义(P>0.05)。结论芬太尼主要在肝脏代谢,无肝期小肠CYP3A1mRNA表达上调,酶活性增加,可能为无肝期芬太尼肝外代谢的机制之一。Objective To investigate the variation of fentanyl concentration and the gene expression and activity of rat intestinal cytochrome P450 3A1 in anhepatic phase. Methods The experiment was comprised of 2 steps. Step 1 ： The rats were randomly divided into experimental group （ group A2, underwent occlusion of the hepatic portal） and control group （group A1 ） , with 10 rats in each group. Fentanyl blood concentration was analyzed by LC/MS/MS. Step 2 ： The rats were randomly divided into group B1 （control） , group B2 and group B3 （the rats underwent devascularization of the hepatic portal for 30 or 60 min）. The levels of CYP3A1 in rat small intestine were assessed with RT-PCR and the enzymic activity of CYP3A1 was detected by fluorometry. Results Fentanyl concentration in anhepatic phase dropped more slowly in group A2 than group A1 （P 〈0.05）, but still remained descending. The gene expression and the enzymic activity of CYP3A1 in rat small intestine were significantly higher in group B2 and group B3 than those in group B1 （P 〈 0.05）. The gene expression of CYP3A1 in rat small intestine between group B2 and group B3 was not significantly different. Conclusion Fentanyl metabolism is mainly in the liver. Higher gene expression and enzymic activity of intestinal CYP3A1 occurs in rats in anhepatic phase than the rats whose hepatic portal is occluded, which may be one of the possible mechanisms for extrahepatic metabolism of fentanyl in anhepatic phase.

关 键 词：芬太尼 细胞色素P450酶 无肝期 血药浓度 RT—PCR 荧光比色法 

分 类 号：R657.3[医药卫生—外科学] R966[医药卫生—临床医学]