摘要
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 and0.01),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.
作者
李晗
张广平
杨依霏
张海静
马梦
陈腾飞
叶祖光
LI Han;ZHANG Guangping;YANG Yifei;ZHANG Haijing;MAMeng;CHEN Tengfei;YE Zuguang(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)
出处
《药物评价研究》
CAS
2020年第2期199-205,共7页
Drug Evaluation Research
基金
国家重点研发计划资助(2018YFC1708204).
