大鼠体内药物代谢酶CYP2C9活性和蛋白表达的测定  

Determination of the activity and protein expression of CYP2C9 in rats

作　　者：高翔[1] 袁明[1] 李永平[1] 朱俊博[1] 尹月淼[1] 李永芳[1] 李向阳[1] 

机构地区：[1]青海大学医学院,青海西宁810001

出　　处：《时珍国医国药》2013年第6期1311-1312,共2页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金资助项目(No.30960457);教育部新世纪优秀人才支持计划(No.NCET-10-0917)

摘　　要：目的测定大鼠体内药物代谢酶CYP2C9的活性和蛋白表达。方法大鼠灌胃给予探针药物苯妥英钠(PHT),采用RP-HPLC法测定给药后12 h血清中苯妥英钠和代谢产物4'-羟基苯妥英(HPPH)的血药浓度,以HPPH与PHT的浓度比值评价CYP2C9的活性。采用ELISA方法测定CYP2C9的蛋白表达。结果 4'-羟基苯妥英和苯妥英钠均在1～20μg.ml-1范围内线性关系良好,低中高3种浓度4'-羟基苯妥英和苯妥英钠的平均回收率分别为101.11%,98.24%,104.32%和96.37%,108.69%,103.54%;平均日内RSD分别为8.10%,4.74%,2.75%和8.40%,3.64%,2.58%;平均日间RSD分别为8.86%,7.36%,5.19%和8.74%,6.33%,8.39%。CYP2C9蛋白定量回归方程为C=98.53×OD-0.86,r=0.999 0。结论建立的方法简便、准确、重现性好,可用于药物代谢酶CYP2C9的活性和蛋白测定。Objective To determine the activity and protein expression of CYP2C9 in rats.Methods After rats was administrated orally phenytoin sodium（PHT）,the serum concentration of PHT and metabolite 4＇-hydroxyphenytoin（HPPH） at 12 h after study drug administration was determined by RP-HPLC.The activity of CYP2C9 was evaluated by the ratio of HPPH to PHT.The protein expression of CYP2C9 was determined by ELISA method.Results A good linearity was obtained in the range of 1 ～ 20 μg.ml-1,and the average recovery of HPPH and PHT was 101.11%,98.24%,104.32% and 96.37%,108.69%,103.54%,respectively.The average value of inter-day RSD was 8.10%,4.74%,2.75% and 8.40%,3.64%,2.58%,respectively,the average value of intra-day RSD was 8.86%,7.36%,5.19% and 8.74%,6.33%,8.39%.The regression equation of CYP2C9 protein determination was C = 98.53 × OD-0.86（r = 0.999 0）.Conclusion The result demonstrated that the method was sensitive,accurate and simple,and can be used to determinate the activity and protein expression of CYP2C9 in rats.

关 键 词：大鼠 CYP2C9 活性 蛋白表达 

分 类 号：R394[医药卫生—医学遗传学]