目的 考察乌头碱配伍人参皂苷Rb₁、甘草苷后对HepG2药物代谢酶（Cytochrome P450，CYP450）中3A4亚型的报告基因荧光活性、mRNA转录及蛋白翻译水平的影响。方法 将pGLuc-CYP3A4报告基因质粒与pcDNA3.1-hPXR表达质粒共转染HepG2细胞，检测乌头类生物碱、人参皂苷和甘草的单体成分对CYP3A4的激活效应；并利用实时荧光定量PCR（qRT-PCR）及Western blotting技术检测人参皂苷Rb₁、甘草苷与乌头碱对CYP3A4 mRNA及蛋白水平的影响。结果 报告基因模型检测结果显示，与对照组比较，乌头碱、新乌头碱、次乌头碱和乙酰乌头碱能下调CYP3A4报告基因荧光强度（P<0.05），其中乌头碱下调能力最强，人参皂苷Rb₁、Rc、Re、Rg1以及甘草苷、异甘草苷、甘草素和甘草酸均能上调报告基因的荧光强度（P<0.05、0.01），其中人参皂苷Rb₁和甘草苷上调能力最明显；同时乌头碱能下调CYP3A4 mRNA与蛋白表达水平（P<0.05、0.01），人参皂苷Rb₁和甘草苷能逆转乌头碱下调CYP3A4的能力（P<0.05、0.01）。结论 人参皂苷Rb₁、甘草苷与乌头碱配伍后可上调CYP3A4的表达，减少乌头碱在体内蓄积时间，起到减毒的作用。

Objective To investigate the effects of aconitine combined with ginsenoside Rb₁ and liquiritin on the fluorescence activity, transcription level and protein translation level of 3A4 subtype of hepatocyte cytochrome P450 (CYP450) in HepG2. Methods The pGLuc-CYP3A4 reporter gene plasmid and pcDNA3.1-hPXR expression plasmid were co-transfected into HepG2 cells to detect the activation effect of aconitines, ginsenoside and components of Glycyrrhiza uralensis on CYP3A4. The expression of ginsenoside Rb₁, liquiritin and aconitine was detected by qRT-PCR and Western blotting. Resluts The results of reporter gene model test showed that aconitine, mesaconitine, hypaconitine and aconitine 3-acetate could down-regulate the fluorescence activity of CYP3A4 reporter gene (P<0.05). Aconitine had the strongest down-regulation ability. Ginsenoside Rb₁, Rc, Re, Rg1 and liquiritin, iso liquiritin, liquiritigenin and glycyrrhiza acid could up-regulate the fluorescence activity of reporter gene (P<0.05 and 0.01), among which ginsenoside Rb₁ and liquiritin were more up-regulation. At the same time, aconitine can down-regulate the expression of CYP3A4 and ginsenoside Rb₁ and liquiritin can reverse the ability of aconitine to down-regulate CYP3A4 (P<0.05, 0.01). Conclusion Ginsenoside Rb₁, liquiritin and aconitine can up-regulate the expression of CYP3A4, reduce the accumulation time of aconitine in vivo and play a role in reducing toxicity.

R285.5

国家重点研发计划资助（2018YFC1708204）