目的 建立肝微粒体测定法,观察中药制剂对CYP3A4 亚型的作用.方法 取大鼠肝脏,制备肝微粒体,分别优化肝微粒体体外温孵系统的反应时间、蛋白浓度及探针药物咪达唑仑的浓度;高效液相色谱-质谱联用(HPLC-MS-MS)法测定大鼠肝微粒体中咪达唑仑的浓度,计算咪达唑仑的活性;在肝微粒体体外温孵系统中分别加入不同浓度的血脂康胶囊、通心络胶囊、枣仁安神胶囊内容物及对照药物酮康唑,测定其半数抑制浓度(IC50)及抑制常数(Ki).结果 肝微粒体体外温孵系统的反应条件为0.4 g·L-1大鼠肝微粒体,4 μmol·L-1咪达唑仑溶液,37 ℃温育5 min.酮康唑、血脂康胶囊、通心络胶囊的IC50分别为(3.4±0.2),(25.1±0.3),(56.2±0.3) mg·L-1,枣仁安神胶囊的IC50﹥150 mg·L-1;酮康唑、血脂康胶囊、通心络胶囊的Ki分别为(1.7±0.1),(25.0±0.2),(50.0±0.3) mg·L-1.结论 建立了中药制剂对CYP3A4亚型作用的大鼠肝微粒体研究模型.血脂康胶囊和通心络胶囊对大鼠CYP3A4亚型有较弱的抑制作用.%Objective To investigate the influence of traditional Chinese medicine preparations on rat liver CYP 3A4 activity in vitro. Methods The rat liver microsomes were prepared from rat livers. The reaction time, protein content and midazolam concentration in the microsomal incubation was optimalized respectively ; the metabolism of midazolam and activity were measured by HPLC-MS-MS;IC50 and Ki of ketoconazole , xuezhikang ,tongxinluo and zaoren ansheng capsule in microsomal incubated system were determined after incubation with the midazolam ( CYP3A4 ). Results The optimum protein content of rat liver microsomes for kinetic analysis was 0.4 g·L-1 with 4 [junol· L midazolam at 37 ℃ for incubation up to 5 min. The IC50 of ketoconazole, xuezhikang and tongxinluo was (3.4±0.2) ,(25.l±0.3) , (56.2±0.3) mg» L ,respectively,but the IC50 of zaoren ansheng capsule was greater than 150 mg · L ; The Ki of ketoconazole, xuezhikang and tongxinluo was ( 1.7 ±0.1) , (25.0 ±0.2),(50.0 ±0.3) mg · L , respectively. Conclusion The model of liver microsomes for evaluation of the influence of traditional Chinese medicine preparations on CYP 3A4 activity is established. xuezhikang and tongxinluo have weak inhibitory effect on rat CYP3A4 in vitro.
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