氟他胺在大鼠肝微粒体经细胞色素P450 1A2代谢的性别差异  

SEX-DIFFERENCE ON FLUTAMIDE METABOLISM IN RAT LIVER MICROSOMAL CYTOCHROME P450 1A2

作　　者：王海学[1] 李端[1] 许长江[1] 刘骁[1] 

机构地区：[1]复旦大学药学院药理学教研室,上海200032

出　　处：《药学学报》2002年第8期608-610,共3页Acta Pharmaceutica Sinica

基　　金：复旦大学创新基金 (CQF3 0 3 80 1) .

摘　　要：目的　体外研究大鼠肝微粒体细胞色素P4 5 0 1A2 (CYP1A2 )对氟他胺 (flutamideFlu)代谢的性别差异影响。方法　制备正常♀♂大鼠肝微粒体 ,用CYP1A2抗体与氟他胺 (2mg·L- 1 )共同温孵 ,测定氟他胺主要代谢产物 2 羟基氟他胺 (2 hydroxyflutamide ,HF)和原药的浓度比 (HF Flu) ,评价氟他胺在大鼠肝微粒体代谢的性别差异。结果　在CYP1A2抗体浓度为 1∶4 0 0 ,孵育时间为 30min条件下 ,氟他胺在♂大鼠肝微粒体中的HF Flu为 (1 5± 0 6 ) ,而♀动物为 (0 9± 0 4 )。不同性别大鼠肝微粒体对氟他胺的代谢存在性别差异 (P <0 0 1)。结论　Flu在♂大鼠肝微粒体中代谢快 ,而在♀大鼠肝微粒体中代谢较慢。AIM To assess the sex difference on flutamide metabolism in rat liver microsomes useing rat cytochrome P450 1A2, inhibitory monoclonal antibody. METHODS Liver microsomes were prepared from male or female rats. Protein concentration and total cytochrome P450 content were determined. Incubation mixture included liver microsomes (1 0 nmol·L -1 ), reduced form of nicotinamide adenine dinucleotide phosphate (NADPH, 0 1 nmol·L -1 ), CYP1A2 (1∶400) and flutamide (2 mg·L -1 ). The incubation time was 30 min. The concentration of flutamide and its major metabolite 2 hydroxyflutamide were analyzed by reverse high performance liquid chromatography. The mobile phase was a mixture of methanol acetonitrile water diethylether (40∶20∶35∶1) with methyltestosterone as internal standard. The detection wavelength was 234 nm. The reaction mixture was extracted with acetic ether 4 mL. Sex difference on flutamide metabolism was expressed as the difference between the concentration ratio of 2 hydroxyflutamide to flutamide in male and female rat liver microsomes. RESULTS The recoveries of flutamide and 2 hydroxyflutamide for the proposed method were more than 75%. The formation of 2 hydroxyflutamide from flutamide was inhibited by CYP1A2 antibodies (1∶400) in male and female rat liver microsome for 30 min of incubation time, but the inhibition of flutamide metabolism in female rat was stronger than that in male. The concentration ratios of 2 hydroxyflutamide to flutamide were (1 5±0 6) and (0 9±0 4) in male and female rat liver microsomes, respectively ( P <0 01). CONCLUSION The results indicate that the activity of male rat CYP1A2 is higher than that of the female rat. There is difference in sex related rate of flutamide metabolism in rat liver microsomes.

关 键 词：氟他胺 2-羟基氟他胺 细胞色素P450 1A2 肝微粒体 

分 类 号：R963[医药卫生—微生物与生化药学]