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目的探讨Rho相关的卷曲蛋白激酶(ROCK)参与乙醇诱导的肠上皮细胞屏障通透性增加的作用。方法应用ROCK特异性抑制剂Y-27632预处理Caco-2细胞后,测定跨上皮细胞阻抗值(TEER)及荧光黄透过率,分析肠上皮细胞屏障的通透性的变化。应用Western blot的方法检测肌球蛋白轻链(MLC)、p-MLC及紧密连接相关蛋白(occludin)的表达。结果 Y-27632显著抑制乙醇诱导的TEER值降低、荧光黄透过率升高、pMLC/MLC及occludin(S/IS)比值的增加。结论 ROCK参与乙醇诱导肠上皮细胞屏障通透性增加。
Objective To investigate the role of Rho-related FOCK protein kinase (ROCK) in the ethanol-induced increase of intestinal epithelial barrier permeability. Methods Caco-2 cells were pretreated with ROCK-specific inhibitor Y-27632, and the transmembrane epithelial cell impedance (TEER) and fluorescence yellow transmittance were measured. The permeability of the intestinal epithelial cell barrier was analyzed. Western blot was used to detect the expression of myosin light chain (MLC), p-MLC and occludin. Results Y-27632 significantly inhibited the decrease of ethanol-induced TEER, the increase of fluorescence yellow transmittance and the increase of pMLC / MLC and occludin (S / IS) ratio. Conclusion ROCK participates in ethanol-induced increase of intestinal epithelial cell barrier permeability.