人原代胎盘滋养层细胞摄取转运模型的建立及氟西汀对胎盘P-gp转运蛋白功能的影响

目的:建立人胎盘滋养层细胞(以下简称"滋养层细胞")转运模型,应用该模型考察氟西汀(fluoxetine,FXT)暴露下外排转运蛋白P-糖蛋白(P-glycoprotein,P-gp)的功能变化,探讨FXT对胎盘转运能力的影响.方法:从足月胎盘中分离、消化、培养得到合胞体滋养层细胞,并从形态学和免疫组织化学等方面对细胞...

Establishment of human placental trophoblasts cell transport model and influence of fluoxetine on P-gp function in placenta

AIM:To establish a placental transport model with primary human placental trophoblasts(hereinafter referred to as "trophoblast cells")and investigate the influence of fluoxetine(FXT)on the function of P-glycoprotein(P-gp)during pregnancy and the transporter involved.METHODS:The primary human placenta trophoblast cells were isolated and validated by morphological observation and immunocytochemical staining. Rhodamine 123,the specific substrates were used to assess the functions of P-gp and to find out the optimal concentrations of substrate and inhibitor. A serials concentration of FXT were incubated with trophoblast cells for 0. 5 h,48 h and 72 h and then the function of P-gp and BCRP were assessed with specific substrate uptake assay.RESULTS:The function of P-gp was conducted with specific substrate and inhibitor. For P-gp the optimal concentration of substrate was 1 μmol/L and the optimal concentration of inhibitors was 10 μmol/L. The cellular accumulation of Rhodamine123 in trophoblast cells was significantly increased after incubation FXT for 0. 5 h,48 h and 72 h(P<0. 05). FXT significantly inhibited the trans-placental transport of P-gp and BCRP. The protein expression of P-gp was significantly downregulated in the presence of FXT(P<0. 05).CONCLUSION:The exposure of FXT may inhibit the expression and function of P-gp.