Aim To investigate the effect of resveratrol on rifampicin-induced P-gp expression through PXR pathway.Methods Dual-Luciferase reporter assays were performed to detect the activity of MDR1 reporter gene in LS174T cells.Real-Time PCR and Western blot were used to detect mRNA and protein expression of P-gp.In addition, rhodamine 123 ( Rh123 ) accumulation assay was used to investigate the P-gp activity.Results Resveratrol ( RES ) could significantly down regulate the rifampicin-induced MDR1 reporter activation both in concentration of 25 and 50 μmol· L-1( P <0.01 ).Besides, RES could also significantly down regulate rifampicin-induced P-gp mRNA expression only in hPXR-transfected cells, but P-gp protein expression was decreased both in hPXR-transfected and non-transfected cells.Furthermore, 25 and 50 μmol· L-1 RES significantly increased rifampicin-inhibited Rh123 accumulation from 77.7% to 91.7% and 95.1%( P <0.05 ), Which indicated that RES could reduce the induction of P-gp activity by rifampicin.Conclusion Resveratrol could affect P-gp expression and activity as an antagonist of human PXR.%目的 研究白藜芦醇通过拮抗hPXR对P-gp基因(MDR1)、蛋白表达及活性的影响.方法 在LS174T细胞中,采用瞬时共转染报告基因实验研究白藜芦醇对PXR介导的MDR1的转录调节作用,并进一步应用Real-Time定量PCR和Western blot方法检测白藜芦醇作用24 h后对利福平诱导的P-gp基因和蛋白变化的影响,罗丹明转运实验考察P-gp活性的变化.结果 双荧光素酶报告基因检测结果显示,25和50 μmol·L-1白藜芦醇可通过拮抗PXR将利福平对MDR1的诱导作用由4.70倍分别降至1.76倍和0.69倍(P<0.01),在过表达hPXR的LS174T细胞中,50 μmol·L-1白藜芦醇可以将利福平诱导的MDR1 mRNA水平由1.8倍降至1.3倍(P<0.05),Western blot结果表明白藜芦醇也可降低利福平诱导的P-gp表达.此外,罗丹明转运实验显示,25和50 μmol·L-1白藜芦醇可以将利福平抑制的累积量由77.7%升至91.7%和95.1%(P<0.05),表明白藜芦醇可降低利福平诱导的P-gp活性.结论 白藜芦醇可以通过拮抗PXR而影响P-gp的基因、蛋白表达及活性.
展开▼
