芍药苷-6-O'-苯磺酸酯对Caco-2细胞中P-gp表达和功能的调控

Regulation of paeoniflorin-6-O'-benzene sulfonate(CP-25)on expression and function of P-glycoprotein in Caco-2 cells

目的探讨芍药苷-6-O'-苯磺酸酯(CP-25)对Caco-2细胞中P-糖蛋白(P-gp)表达和功能的调控作用。方法体外培养人结肠癌细胞Caco-2,以表观渗透系数(Papp)为指标,研究pH对CP-25转运的影响;流式细胞术检测CP-25对P-gp的表达和活性的影响;Western blot法检测Caco-2细胞中的Phospho-NF-кB p65和P-gp表达;激光共聚焦法检测Caco-2细胞中Phospho-NF-кB p65的表达。结果Papp值表明CP-25在结肠中的吸收高于Pae;流式细胞术检测结果显示,CP-25可增加细胞内RHO 123的积累,下调P-gp表达;Western blot结果显示CP-25作用后Phospho-NF-кB p65和P-gp的表达下调;激光共聚焦检测结果显示CP-25可使Phospho-NF-кB p65表达下调。结论CP-25能够下调Caco-2细胞中P-gp的表达,抑制其转运活性,其作用机制可能与阻断NF-кB信号通路激活有关。

Objective To study the regulation of CP-25 on P-glycoprotein(P-gp)expression and function in Caco-2 cells.Methods Human colon cancer cell line Caco-2 was cultured in vitro.The apparent permeability coefficient(Papp)was used as an indicator to study the effect of pH on the transport of CP-25.Flow cytometry was used to detect the expression and activity of CP-25 on P-gp.The expression of Phospho-NF-кB p65 and P-gp in Caco-2 cells was detected by Western blot.The expression of Phospho-NF-кB p65 in Caco-2 cells was detected by laser confocal microscopy.Results The Papp value indicated that CP-25 was higher in the colon than in Pae.The results of flow cytometry showed that CP-25 significantly increased the accumulation of RHO123 in cells and down-regulated the expression of P-gp.Western blot results showed that CP-25 could significantly down-regulate the expression of Phospho-NF-кB p65 and P-gp after treatment.Laser confocal assay showed that CP-25 could down-regulate the expression of Phospho-NF-кB p65.Conclusion CP-25 can down-regulate the expression of P-gp in Caco-2 cells and inhibit its transport activity,which may be related to the activation of NF-кB signaling pathway.