七叶皂苷对P-糖蛋白功能的影响

Effect of aescin on P-glycoprotein activity

目的:探讨七叶皂苷对P-糖蛋白功能的影响。方法:构建稳定表达P-糖蛋白的LLC-PK1细胞系,以real-time RT-PCR和Western Blotting分析P-糖蛋白基因mRNA和蛋白表达,共聚焦显微镜观察P-糖蛋白细胞定位,流式细胞术检测细胞内罗丹明123荧光强度。结果:(1)P-糖蛋白在LLC-PK1细胞中稳定高表达;(2)转染细胞中P-糖蛋白定位在细胞膜上;(3)七叶皂苷抑制P-糖蛋白功能,细胞内罗丹明123荧光强度增加123%,但其抑制效果是维拉帕米的30%。结论:七叶皂苷抑制P-糖蛋白功能,但其抑制效果弱于维拉帕米。

Objective：This study was to investigate the effect of aescin on P-glycoprotein（P-gp） activity.Methods：MDR1 was transfected into LLC-PK1 cells.Expressions of mRNA and protein of MDR1 were measured by real-rime RT-PCR and Western Blotting,respectively,localization of P-gp by confocal microscopy.The fluorescence intensity of Rho123 was assayed by flow cytometry（FCM）.Results：（1） Stable transfection MDR1 could increase the P-gp expression in RNA and protein level;（2） P-gp located in the membrance of LLC-PK1 cells;（3） Aescin could inhibit the activity of P-gp and enhance 123% intracellular fluorescence intensity of Rho123,but the effect inhibition was 30% that of verapamil.Conclusion：Aescin could inhibit P-gp activity,but the effect was weaker than verapamil.