Tan Jia Cheng, Cui Wen Xia, Heng Ding, Lin Lin
Department of Gastroenterology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu Province, China.
J Dig Dis. 2014 Aug;15(8):409-18. doi: 10.1111/1751-2980.12163.
To investigate the alterations of esophageal epithelial barrier during the process of reflux esophagitis (RE).
In total, 85 Sprague-Dawley rats were randomly divided into two groups, the sham-operation group (n = 25) and the RE group induced by incomplete pyloric ligation (n = 60). The establishment of RE model and the severity of esophagitis were evaluated by hematoxylin and eosin stain. Dilated intercellular spaces (DIS) in the esophageal epithelium were observed by transmission electron microscopy. The cellular distributions of ZO-1, occludin and claudin-1 were assessed by immunohistochemical stain. The expressions of these tight junction (TJ) proteins and the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), myosin light chain (MLC) and nonmuscular myosin light chain kinase (nmMLCK) were analyzed by Western blot.
DIS occurred gradually in the RE group. ZO-1, occludin and claudin-1 were incompletely or even not expressed in the RE group. TJ proteins were expressed in the membrane instead of the cytoplasm in many epithelial cells in RE. With Western, the expression of ZO-1, occludin and claudin-1 was increased gradually in the RE group (P < 0.05). The phosphorylation levels of nmMLCK, MLC and ERK1/2 were also increased (P < 0.05). There was no marked changes in the esophageal epithelium in the sham-operation group.
TJ proteins could be used as sensitive markers of RE instead of DIS. ERK1/2 may participate in regulating TJ proteins in esophageal epithelia in RE.
探讨反流性食管炎(RE)过程中食管上皮屏障的变化。
将85只Sprague-Dawley大鼠随机分为两组,即假手术组(n = 25)和不完全幽门结扎诱导的RE组(n = 60)。通过苏木精-伊红染色评估RE模型的建立及食管炎的严重程度。用透射电子显微镜观察食管上皮细胞间扩张间隙(DIS)。通过免疫组织化学染色评估紧密连接蛋白1(ZO-1)、闭合蛋白和Claudin-1的细胞分布。采用蛋白质免疫印迹法分析这些紧密连接(TJ)蛋白的表达以及细胞外信号调节激酶1/2(ERK1/2)、肌球蛋白轻链(MLC)和非肌肉型肌球蛋白轻链激酶(nmMLCK)的磷酸化水平。
RE组DIS逐渐出现。RE组中ZO-1、闭合蛋白和Claudin-1表达不完全甚至未表达。在RE组中,许多上皮细胞的TJ蛋白表达于细胞膜而非细胞质。蛋白质免疫印迹法结果显示,RE组中ZO-1、闭合蛋白和Claudin-1的表达逐渐增加(P < 0.05)。nmMLCK、MLC和ERK1/2的磷酸化水平也升高(P < 0.05)。假手术组食管上皮无明显变化。
TJ蛋白可作为RE的敏感标志物,而非DIS。ERK1/2可能参与调节RE中食管上皮的TJ蛋白。