艾滋病中药和化学药通过CYP3A4的代谢性相互作用的体外研究

In vitro herb-drug interaction mediated by CYP3A4 and its potential application in the treatment of AIDS

目的:评价应用于艾滋病治疗的24个中药的提取物对人肝微粒体细胞色素P450氧化代谢酶3A4(CYP3A4)活性的抑制作用,为阐明中药和化学药在治疗艾滋病时的代谢性相互作用的增效机制提供数据。方法:用标准化方法提取24个中药的65个部位,并在生理条件下与混合人肝微粒体进行预孵15 min后,加入探针底物睾酮和辅酶β-NADPH进行孵育30 min,用LC-MS/MS对CYP3A4的活性进行定量检测,计算抑制率(IC50)。结果:在24个中药中虎杖和黄芩对CYP3A4具有较强的抑制,其不同部位的IC50分别为3.25～8.19和10.0～29.0μg.mL-1。其他测试的中药提取物,如甘草、苦参、牡丹皮、土茯苓和乌药,它们的不同提取部位对CYP3A4的抑制作用有较大差异。而败酱草,板蓝根,当归,黄芪和龙胆对CYP3A4几乎不具抑制效果。结论:中药的不同提取物对CYP3A4的抑制效果有较大差异。

Objective： To evaluate CYP3A4-mediated interaction of herb medicines with HIV protease inhibitors, and to explore the potential efficacy enhancement by herb-drug metabolic interaction in the treatment of AIDS. The 24 traditional Chinese medicines （TCM） for AIDS treatment and the HIV protease inhibitor indinavir were studied. Methods： Totally 65 extracts from 24 TCM isolated by a standardized method were pre2incubated with pooled human liver microsomes for 15 min, and then incubated with β-NADPH and testosterone as CYP3A4 probe substrate for 30 min. The enzyme activity of CYP3A4 was measured by LC-MS/MS and the IC50 of six extracts were calculated. Results： The extracts from Rhizoma Et Radix Polygoni Cuspidati and Radix Scutellariae showed stronger inhibitory effects on CYP3A4 with IC50 3.25 - 8.19 and 10.0 - 29.0 μg· mL^-1, respectively, among the 24 test herbs. The extracts from Radix Et Rhizoma Glycyrrhizae, Radix Sophorae Flavescentis, Cortex Moutan, Rhizoma Smilacis Glabrae and Radix Linderae showed different inhibitory effects. No significant inhibition on CYP3A4 was observed in the extracts from Herba Patriniae, Radix Isatidis, Radix Angelicae Sinensis, Radix Astragali and Radix Et Rhizoma Gentianae. Conclusion： The inhibitive effect of different extracts from TCM on enzyme activity of CYP3A4 was much different.