建立用大鼠肝微粒体快速考察药物体外CYP3A1活性抑制的方法  

Establish A Method to Evaluate Activity of Rat Liver Microsomal CYP3A1 that Inhibited in vitro

作　　者：梁蔚婷[1] 黄红兵[1] 刘韬[1] 

机构地区：[1]华南肿瘤学国家重点实验室中山大学肿瘤防治中心药学部,广东广州510060

出　　处：《今日药学》2012年第1期13-16,共4页Pharmacy Today

基　　金：广东省医院药学研究基金资助项目(编号:2008A007);广东省科技计划项目(编号:2009B030801160)

摘　　要：目的研究酮康唑对大鼠肝微粒体细胞色素P450同工酶3A1(CYP3A1)活性的作用,建立一种用大鼠肝微粒体快速考察药物体外CYP3A1活性抑制的方法。方法采集大鼠肝微粒体,随机分为对照组和药物处理组。酮康唑药物处理组分别加入不同浓度的酮康唑,对照组只加入培养液,混匀后放入37℃培养箱中孵育15 min,然后加入CYP 3A1底物睾酮,继续孵育10 min,最后加入内标物氢化可的松。用HPLC法测定睾酮经大鼠肝微粒体温孵后生成6-β羟基睾酮的量,代表CYP3A1的活性。结果各个处理组与对照组的比较差异均有显著性差异(P<0.05);提示在一定浓度范围内,大鼠同工酶CYP3A1的相对活性百分比随着酮康唑浓度的增加而逐渐减低,各处理组与对照组比较差异均有显著性意义(P<0.5)。半数抑制浓度(IC50)值为3.25μg/ml。结论酮康唑对大鼠肝微粒体细胞色素P450同工酶3A1的活性有强抑制作用,建立的通过大鼠肝微粒体快速考察药物体外CYP3A1探针抑制的方法重复性和稳定性均良好,为评价新药对大鼠肝微粒体CYP3A1的活性的体外作用提供可靠的检测手段。Objective To study the effect of ketoconazole on the activity of rat liver microsomal cytochrome P450 isoenzyme 3A1（CYP 3A1）,and to establish a method to evaluate the activity of CYP3A1 that inhibited in vitro.Methods Rat liver microsomes were obtained and randomly divided into control group and ketoconazole-treatment groups at different concentrations.After 15 min of culture at 37 ℃,testosterone were added in the substrates,and incubated for another 20 min with hydrocortisone finally added as internal standard.The metabolites 6β-hydroxytestosterone were then measured with high performance liquid chromatography to access the activity of CYP 3A1.Results Significant differences in the relative activity of CYP 3A1 were found between the treatment and the control groups（P〈0.05）,showing that ketoconazole could inhibit CYP450 3A1 activity in a concentration-dependent fashion markedly.The 50% inhibiting concentration（IC50） of ketoconazole for CYP3A4 was 3.25 μg/ml.Conclusion Ketoconazole can strongly inhibit the activity of CYP450 3A1 in liver microsome of healthy rats,the method that evaluates the activity of CYP3A1 that inhibited in vitro is reliable,convenient and more efficient and can be applied to evaluate the NHE＇s activity of CYP3A1 in vitro.

关 键 词：酮康唑 肝微粒体 细胞色素P450 CYP3A1 CYP3A4 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]