Effect of miR-200b on intestinal epithelial tight junction via MLCK/P-MLC signaling pathway
10.3969/j.issn.1000-3606.2016.07.015
- VernacularTitle:miR-200b调控肌球蛋白轻链激酶/磷酸化肌球蛋白轻链信号通路对肠上皮紧密连接的影响
- Author:
Yujie SHEN
;
Cong ZHANG
;
Yingwei CHEN
- Publication Type:Journal Article
- Keywords:
miR-200 b;
intestinal epithelial tight junction;
myosin light chain kinase;
phosphorylated myosin light chain;
cell
- From:
Journal of Clinical Pediatrics
2016;34(7):540-543
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the impact and mechanism of miR-200 b on intestinal epithelial tight junction. Methods The negative-lentivirus and human-miR-200 b-lentivirus were employed to infect the Caco-2 cell thus establishing two stable cell lines which were then stimulated by 10 ng/mL human tumor necrosis factor-α(TNF-α) to establish the model of the intestinal epithelial injury. Those Caco-2 cells were divided into NC, NC+TNF-α, 200b, and 200b+TNF-αgroups.The tight junction permeability was detected by transepithelial electrical resistance (TEER) and Fluorescein isothiocyanate-labeled dextran (FITC-dextran). The protein alterations myosin light chain kinase (MLCK)/phosphorylated myosin light chain (P-MLC) pathways were measured by Western blot analysis. Results Compared to NC group, NC+TNF-αgroup had lower TEER, higher FITC-dextran, and up-regulated expressions of MLCK and P-MLC proteins (P0 . 05 ). Compared to NC+TNF-αgroup, 200 b+TNF-αgroup had higher TEER, lower FITC-dextran and down-regulated expressions of MLCK and P-MLC proteins (P<0 . 05 ). Conclusion miR-200 b ameliorated TNF-α-induced intestinal epithelial tight junction disruption via regulation MLCK/P-MLC pathway.