CYP3A5基因型的差异对环孢素药代动力学影响的体外研究  

Effects of CYP3A5 genotype on CsA pharmacokinetics variances in vitro

作　　者：张澈[1] 涂自良[1] 胡永芳[2] 王启斌[1] 程晓丽[1] 张蓬华[1] 周宏灏[2] 

机构地区：[1]郧阳医学院附属太和医院药学部,湖北十堰442000 [2]中南大学临床药理学研究所,湖南长沙410083

出　　处：《中国药理学通报》2007年第9期1131-1137,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30371668)

摘　　要：目的体外明确CYP3A5基因多态性对CYP3A5蛋白酶表达量的影响;不同的CYP3A5基因型对环孢素体外代谢的影响。方法采用PCR-RFLP方法和测序分析了21份肝脏标本的CYP3A5*3基因型;Western-blot方法对蛋白质进行定量;将CYP3A4特异性抑制剂酮康唑和环孢素一起孵育后,采用LC/MS-MS检测AM9/CsA浓度。结果CYP3A5*1/*1、CYP3A5*1/*3基因型和CYP3A5*3/*3基因型样本的蛋白表达量分别为:260.10±136.89nmol.g-1,85.44±55.22nmol.g-1和19.88±17.33nmol.g-1。各基因型蛋白表达量存在差异的同时,也存在一定程度的性别差异。采用AM9/CsA来评估CYP3A5活性,人肝微粒体孵育环孢素,CYP3A5*1/*1的活性明显强于CYP3A5*3/*3;量化校正CYP3A5后,此活性不存在性别差异。结论CYP3A5基因多态性可引起蛋白酶表达量的差异,因而对携带有CYP3A5*3个体,应用比CYP3A5*1/*1个体更小剂量的环孢素就能达到所需要的治疗血药浓度。Aim To identify the association between CYP3A5 genotype and CYP3A5 protein and to evaluate the influence the CYP3A5 gene polymorphisms on CsA metabolism in vitro. Methods Through polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） and sequencing assays, the allelic frequency of the CYP3A5^ ＊ 3 of 21 liver samples was determined. CYP3A5 protein quantity was determined by western-blot; After liver microsomes incubation with CsA and ketoconazole, the AM9 and CsA concentration was detected by LC/MS-MS. Results The protein conceutration of CYP3A5^ ＊ 1/＊ ^1 ,CYP3A5 ^＊ 1/＊^ 3 and CYP3A5^ ＊ 3/＊^ 3 genotype were （ 260. 10 ± 136. 89 ） nmol·g^-1, 85.44 ± 55.22 nmol·g^-1 and 19. 88 ± 17.33 nmol·g^-1, respectively. Different genotype had different protein expression, but there was not stadistical difference between sexeven in the same genotype. CsA incubation in vitro suggested the enzyme activity of HLM from CYP3A5^ ＊ 1/＊^ 1 genotype sample was much higher than that from CYP3A5^ ＊ 3 genotype sample when using AM9/CsA as the appraisal index. Conclusion The polymorphisms of CYP3A5 might induce the quantity difference of CYP3A5 protein. Compared with CYP3A5 ^＊ 1/＊ ^1 individual, ones with CYP3A5 ^＊^3 allele would need a fewer dosage of CsA to obtain the target concentration.

关 键 词：CYP3A5 肝微粒体孵育 环孢素 

分 类 号：R322.47[医药卫生—人体解剖和组织胚胎学] R345.99[医药卫生—基础医学]