大鼠体内药物代谢酶CYP2C19活性和蛋白表达的测定方法建立  

ESTABLISHMENT OF DETERMINATION METHOD OF THE ACTIVITY AND PROTEIN EXPRESSION OF CYP2C19 IN RATS

作　　者：王学军[1] 袁明[2] 李永芳[2] 朱俊博[2] 李永平[2] 李向阳[2] 

机构地区：[1]青海红十字医院 [2]青海大学医学院

出　　处：《青海医学院学报》2012年第4期260-263,共4页Journal of Qinghai Medical College

基　　金：国家自然科学基金资助项目(30960457);教育部新世纪优秀人才支持计划(NCET-10-0917)

摘　　要：目的建立大鼠体内药物代谢酶CYP2C19的活性和蛋白表达的测定方法。方法大鼠灌胃给予探针药物奥美拉唑(OMZ),采用RP-HPLC法测定给药后3 h血清中奥美拉唑和代谢产物5-羟基奥美拉唑(5-OHOMZ)的血药浓度,以5-OHOMZ与OMZ的浓度比值评价CYP2C19的活性。采用ELISA法测定CYP2C19的蛋白表达。结果 OMZ和代谢产物5-OHOMZ分别在1～20μg.mL-1和0.5～12μg.mL-1范围内线性关系良好,低、中、高3种浓度OMZ和5-OHOMZ的平均回收率分别为88.55%、93.50%、92.91%和89.04%、97.59%、89.50%;平均日内RSD分别为9.92%、7.92%、6.29%和5.84%、7.49%、3.59%;平均日间RSD分别为8.68%、8.79%、8.02%和9.14%、7.99%、7.80%。CYP2C19蛋白定量回归方程为C=108.04×OD-29.41(r=0.9956)。结论建立的方法简便、准确、重现性好,可用于药物代谢酶CYP2C19的活性和蛋白测定。Objective To establish the determination method of the activity and protein expression of CYP2C19 in rats.Methods After rats was administrated orally omeprazole（OMZ）,The serum concentration of OMZ and metabolite 5-hydroxy omeprazole（5-OHOMZ）at 3 h after study drug administration was determined by RP-HPLC.The activity of CYP2C19 was evaluated by the ratio of 5-OHOMZ to OMZ.The protein expression of CYP2C19 was determined by ELISA method.Results A good linearity was obtained in the range of 1-20 μg·mL-1 and 1-20 μg·mL-1 for omeprazole and 5-hydroxy omeprazole,respectively,and the average recovery of OMZ and 5-OHOMZ was 88.55%,93.50%,92.91% and 89.04%,97.59%,89.50%,respectively.The average value of inter-day RSD was 9.92%,7.92%,6.29% and 5.84%,7.49%,3.59% respectively,The average value of intra-day RSD was 8.68%,8.79%,8.02% and 9.14%,7.99%,7.80%.The regression equation of CYP2C19 protein determination was C= 108.04×OD-29.41（r= 0.9956）.Conclusion The result demonstrated that the method was sensitive,accurate and simple,and can be used to determination the activity and protein expression of CYP2C19 in rats.

关 键 词：大鼠 CYP2C19 活性 蛋白表达 测定 

分 类 号：R91[医药卫生—药学] R730.53