高浓度胆汁改变肠上皮细胞氯通道蛋白－2和通透性

目的 观察胆汁对肠上皮细胞氯离子通道和通透性的影响.方法 鼠肠上皮细胞株IEC-6分别与5.0％、1.0％、0.1％胆汁及氯通道激动剂接触.20h后检测跨膜电阻,Western blot分析氯离子通道蛋白-2(CLC-2)和紧密连接闭锁小带－1(ZO-1)表达的变化及各条带的相对灰度值.结果 5.0％浓度组降低跨膜电阻作用最强.5.0％和1.0％组的CLC-2蛋白相对灰度值(0.30±0.05和0.37±0.08)低于对照组(P均＜0.05).5.0％组的ZO-1相对表达量下降.添加氯通道激动剂后,5.0％浓度组的跨膜电阻(451.3±60.5)Ω.cm2及ZO-1相对表达量(0.32±0.04)较对照组上升明显(P均＜0.05).结论 高浓度胆汁破坏肠上皮细胞氯离子通道和紧密连接蛋白,增加上皮细胞通透性.

High concentration bile changes the chloride channel protein-2 and permeability of intestinal epithelium cell line

Objective To investigate the effects of bile on chloride channel protein-2 (CLC-2)and permeability of intestinal epithelium cell.Methods Rat intestinal epithelium cell line IEC-6 was cultured.They were exposed to 5.0％,1.0％,0.1％ rat bile and chloride channel agonist Lubiprostone.After 20 h cultured,transepithelial electrical resistance (TER) of the monostratal cells was measured.The change of CLC-2 and zonula occludens-1 (ZO-1) were examined by Western blotting,and the images were analysed quantitatively.Results The TEER of 5％ group was most lower.The average relative gradations of Western blotting images in 5.0％ group and 1.0％ were lower obviously than those in control group (0.30 ± 0.05,0.37 ± 0.08 vs.0.56 ± 0.08) ( all P ＜ 0.05 ).The relative gradations of ZO-1 were decreaseded only in 5.0％ group.After Lubiprostone added,the TER in 5.0％ group was upgraded (451.3 ± 60.5 )Ω·cm2,and the average relative gradations of ZO-1 (0.32 ± 0.04).Conclusion High concentration of bile destroied enterocyte chloride channel and tight junction protein,increase epithelium permeabihty.