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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: To investigate the effect of aescine on P-glycoprotein function. METHODS: The LLC-PK1 cell line stably expressing P-glycoprotein was constructed. The mRNA and protein expression of P-glycoprotein were analyzed by real-time RT-PCR and Western Blotting. The localization of P-glycoprotein was observed by confocal microscopy. Intracellular detection of intracellular rhodamine 123 fluorescence intensity. Results: (1) P-glycoprotein was stably overexpressed in LLC-PK1 cells; (2) P-glycoprotein was located on the cell membrane in transfected cells; (3) Aescin inhibited P-glycoprotein function, Rhodamine 123 increased the fluorescence intensity by 123%, but its inhibitory effect was 30% of that of verapamil. Conclusion: Aescin can inhibit P-glycoprotein function, but its inhibitory effect is weaker than that of verapamil.