何首乌6种成分对人孕烷X受体介导的CYP3A4的调控作用

Effect of six components in Polygoni Multiflori Radix on regulation of CYP3A4 mediated by human pregnane X receptor

该文探讨何首乌中6种化学成分(没食子酸、槲皮素、山柰酚、木犀草素、白藜芦醇、芹黄素)对人孕烷X受体(humanpregnant X receptor,PXR)介导的CYP3A4的转录调控作用。利用前期建立的PXR介导的CYP3A4药物诱导快速筛选技术,采用MTS细胞检测方法考察化学成分对细胞活性的影响,计算IC50;利用双荧光素酶报告基因系统,将PXR表达载体和含CYP3A4转录调控区的报告基因载体共转染HepG2细胞,10μmol·L-1利福平为阳性对照,10μmol·L-1酮康唑为阴性对照,用何首乌中6种成分--没食子酸、槲皮素、山柰酚、木犀草素、白藜芦醇、芹黄素(5,10,20μmol·L-1)分别处理24 h,进行双荧光素酶活性检测。结果表明,空载质粒pc DNA3. 1与p GL4. 17-CYP3A4共转染时没食子酸、白藜芦醇对CYP3A4的调控为抑制效应,槲皮素、山柰酚、木犀草素对CYP3A4产生诱导作用;pc DNA3. 14-PXR与p GL4. 17-CYP3A4共转染时,槲皮素、山柰酚、木犀草素、白藜芦醇、芹黄素均产生诱导效应。综上所述,6种成分对CYP3A4产生抑制或激活效应,PXR参与后,5种成分对CYP3A4产生诱导效应,其中3种成分的诱导效应具有显著性差异。该研究提示,在与何首乌联合用药时应注意潜在的药物相互作用,提高安全性和有效性。

This paper aimed to study the six chemical components of Polygoni Multiflori Radix(gallic acid,quercetin,luteolin,kaempferol,resveratrol,apigenin). By the established pregnane X receptor(human pregnant X receptor,PXR) CYP3 A4 mediated drug induced rapid screening technique,the effect of chemical components on the cell activity was detected by MTS cell method,and the value of IC50 was calculated. The dual luciferase reporter system was used to co-transfect PXR reporter gene expression vector containing transcriptional regulation and CYP3 A4 with HepG2 cells,with 10 μmol·L-1 rifampicin(RIF) as a positive control,and 10μmol·L-1 of ketoconazole(TKZ) as negative control. Gallic acid,quercetin,luteolin,kaempferol,apigenin,resveratrol(5,10,20μmol·L-1) were used to incubate for 24 h,and the luciferase activity was detected. The results showed that when plasmid pc DNA3. 1 was co-transfected with p GL4. 17-CYP3 A4,gallic acid and resveratrol had an inhibitory effect on the regulation of CYP3 A4,and quercetin,luteolin,kaempferol had an inductive effect on CYP3 A4;when pc DNA3. 14-PXR was co-transfected with p GL4. 17-CYP3 A4,quercetin,luteolin,kaempferol,apigenin,resveratrol had an inductive effect. To sum up,the 6 reported liver injury components had inhibitory or activating effects on CYP3 A4. After PXR plasmid was involved,5 components had an inductive effect on CYP3 A4,and the inductive effects of 2 components were significantly different. In this experiment,we found that 2 kinds of potential liver injury components in Polygoni Multiflori Radix had been induced by CYP3 A4,which was achieved through PXR regulation. It suggested that attention shall be paid to potential drug interactions when combined with Polygoni Multiflori Radix,so as to improve the safety and efficacy.