摘要
该文探讨何首乌主要化学成分2,3,5,4'-四羟基芪-2-O-β-D-吡喃葡萄糖苷(2,3,5,4'-tetrahydroxystilbene-2-O-β-Dglucopyranoside,THSG)、蒽醌类(大黄素、大黄酸、大黄酚、芦荟大黄素、大黄素甲醚)对人孕烷X受体(human pregnant X receptor,PXR)介导的CYP3 A4的转录调控作用。利用前期建立的PXR介导的CYP3 A4药物诱导快速筛选技术,采用MTS细胞检测方法考察何首乌中主要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CYP3 A4转录调控区的报告基因载体共转染Hep G2细胞,10μmol·L^(-1)利福平(RIF)为阳性对照,10μmol·L–1酮康唑(TKZ)为阴性对照,用THSG(2.5,5,10μmol·L^(-1))和蒽醌类成分(2.5,5,10μmol·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,THSG、大黄酚、大黄素甲醚、大黄酸、芦荟大黄素对CYP3A4的调控多为抑制效应,大黄素对CYP3A4产生诱导作用;pc DNA3.14-PXR与p GL4.17-CYP3A4共转染时,THSG、大黄酚、大黄素甲醚、大黄酸、芦荟大黄素均产生诱导效应。综上,THSG和蒽醌类成分均可对CYP3A4产生抑制或激活效应,PXR参与后,上述成分对CYP3A4均产生诱导效应,提示何首乌中主要成分对CYP3A4的诱导作用是通过PXR实现的。以上结果提示在与何首乌联合用药时应注意潜在的药物相互作用,提高中药的安全性和有效性。
The rapid screening technology was used to investigate the transcriptional regulation effect of main chemical constituents in tubers of Polygonum multiflorum, including 2,3,5,4＇-tetrahydroxystilbene-2-O-β-D-glucopyranoside（THSG） and anthraquinones （such as rhein, chrysophanol, aloe-emodin, emodin） on CYP3A4 drug inducers induced by human pregnancy X receptor （PXR）.The effect of chemical composition on the cell activity was detected by MTS cell viability assay. IC50 was calculated. The expression vector and the reporter vector were co-transfected into HepG2 cells, with 10 μmol·L^-1 rifampicin （RIF） as a positive control, and 10 μmol·L^-1 ketoconazole （TKZ） as a negative control. After treated with different concentrations of anthraquinones （2.5, 5, 10 μmol·L^-1） for 24 h, the cells were tested for dual luciferase activity. The results show that the inhibitory effect of THSG, chrysophanol, emodin, rhein and aloe-emodin on CYP3A4 was inhibited by co-transfection of pcDNA3.1 and pGL4.17-CYP3A4. The expressions of pcDNA3.14-PXR and pGL4.17-CYP3A4 were induced by the four compounds. Besides, emodin had a direct inducing effect. In conclusion, the four anthraquinone compounds have an inducing effect on CYP3A4 by PXR, but emodin can directly induce CYP3A4. THSG can inhibit CYP3A4, but plasmid can induce CYP3A4 after intervened with PXR.These results suggest that we should pay attention to the liver function and avoid liver damage in the combined administration of drugs.
出处
《中国中药杂志》
CAS
CSCD
北大核心
2017年第24期4827-4833,共7页
China Journal of Chinese Materia Medica
基金
北京市自然科学基金项目(7164291)
国家"重大新药创制"科技重大专项(2015ZX09501004-003-003)
中医药行业科研专项(201507004)
作者简介
张照研，博士研究生，研究方向为中药药理毒理，Tel：（010）66930267，E-mail：zhaoyanzhang@emails．bjut.edu．cn;王宇光．副研究员，博士，研究方向为中药药理毒理，Tel：（010）66931225，E—mail：wyg79@163．com;[通信作者]高月，研究员，博士，博士生导师，研究方向为中药药理毒理，Tel：（010）66931312，E-mail：gaoyue@nic．bmi．ac．cn；
