三七总皂苷对细胞色素P450酶CYP3A4和CYP2C9的影响

Effect of Panax Notoginseng Saponins on Cytochrome P450 Enzymes CYP3A4 and CYP2C9

目的探讨三七总皂苷对Chang Liver细胞P450酶(CYP450) CYP3A4和CYP2C9表达的影响。方法采用CCK-8法考察三七总皂苷对Chang Liver细胞活力的影响,确定给药剂量范围;采用定量RT-PCR法考察三七总皂苷在给药后4,8,24,48 h对Chang Liver细胞中CYP3A4和CYP2C9 mRNA表达的影响;通过蛋白质印迹(Western Blot)法考察三七总皂苷在给药后4,8,24,48 h对Chang Liver细胞中CYP3A4和CYP2C9蛋白表达的影响。结果 100μg/m L三七总皂苷干预Chang Liver细胞4,8,24,48 h后,各干预组CYP2C9蛋白表达量明显高于空白对照组(P <0. 05),各组诱导水平随着作用时间的延长相应增加。8 h组、24 h组、48 h组CYP3A4蛋白表达量明显高于空白对照组(P <0. 05),各组的相对蛋白表达量呈时间依赖性,8 h组、24 h组、48 h组CYP2C9和CYP3A4的mRNA表达量均明显高于空白对照组(P <0. 05),且各组的诱导水平随着作用时间的延长相应增加。结论三七总皂苷可诱导CYP3A4和CYP2C9的mRNA及蛋白的表达,临床在使用三七总皂苷时,需要关注其与CYP3A4和CYP2C9底物药物之间可能的相互作用。

Objective To investigate the effect of panax notoginseng saponins(PNS) on the expression of cytochrome P450 enzymes(CYP 450)CYP3A4 and CYP2C9 in Chang Liver cells. Methods The effect of PNS on the viability of Chang Liver cells were investigated by CCK-8 assay,and the dosage range was determined. The effect of PNS on the expression of CYP3A4 and CYP2C9 mRNA in Chang Liver cells were investigated by quantitative RT-PCR method at 4, 8, 24 and 48 h after administration. The effect of PNS on the expression of CYP3A4 and CYP2C9 protien in Chang Liver cells were investigated by Western Blot method at 4,8,24 and 48 h after administration. Results Compared with the blank control group,the expression of CYP2C9 protein in Chang Liver cells was significantly higher after 4,8,24 and 48 h of treatment with 100 μg/mL PNS( P < 0. 05), and the induction level of each group increased with the prolongation of the action time. The expression of CYP3A4 protein in 8 h group,24 h group and 48 h group was significantly higher than that in the blank control group( P < 0. 05),and the relative expression of CYP3A4 protein in each group was time-dependent.The expression of CYP2C9 and CYP3A4 mRNA in 8 h group, 24 h group and 48 h group were significantly higher than that in the blank control group( P < 0. 05), and the induction level of each group increased with the prolongation of action time. Conclusion PNS can induce the expression of CYP3A4 and CYP2C9 mRNA and protien. It is necessary to pay attention to the possible drug-drug interaction between PNS and CYP3A4 and CYP2C9 substrates in clinic.