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目的:研究洛美利嗪对原代培养的大鼠脑微血管内皮细胞(RBMECs)上P-糖蛋白(P-gp)功能和表达的影响。方法:使用流式细胞术分析洛美利嗪对P-gp底物-罗丹明123(rhodamine123,Rh123)在RBMECs中外排的影响,使用流式细胞术分析了洛美利嗪对RBMECs上P-gp表达的影响,应用RT-PCR技术分析了洛美利嗪对RBMECsmdr1基因mRNA水平表达的影响,还使用Transwell模型研究了洛美利嗪对Rh123通透RBMECs单层细胞转运的影响。结果:洛美利嗪通过抑制了RBMECs胞内Rh123的外排;洛美利嗪对P-gp功能的影响与RBMECs上P-gp及mdr1基因mRNA表达无关;在Transwell模型中,洛美利嗪还能显著增加Rh123跨RBMECs单层细胞膜的、经上室至下室的转运,并抑制相反方向的Rh123的转运。结论:洛美利嗪能显著地抑制RB-MECs上P-gp的活性,并对P-gp底物的跨细胞转运产生影响。
AIM: To investigate the effect of lomerizine on the function and expression of P-glycoprotein (P-gp) in primary cultured rat brain microvascular endothelial cells (RBMECs). Methods: Flow cytometry was used to analyze the effect of lomerizine on the efflux of P-gp substrate rhodamine123 (Rh123) in RBMECs. Flow cytometry was used to analyze the effect of lomerizine on RBMECs P- The effect of Lomerizine on mRNA expression of RBMECsmdr1 gene was analyzed by RT-PCR. Transwell model was also used to study the effect of Lomerizine on the monolayer cell transport of Rh123-permeabilized RBMECs. Results: Lomerizine inhibited the efflux of intracellular Rh123 in RBMECs. The effect of Lomerizine on the function of P-gp was not related to the expression of P-gp and mdr1 mRNA in RBMECs. In the Transwell model, It also significantly increased the translocation of Rh123 across the monolayer of RBMECs across the suprachamber to the inferior compartment and inhibited Rh123 transport in the opposite direction. Conclusion: Lomerizine can significantly inhibit the activity of P-gp on RB-MECs and affect the transcellular transport of P-gp substrates.