miR-34a/HSP70通路在肠道成纤维细胞影响克罗恩病术后吻合口纤维化中的作用

项目名称： miR-34a/HSP70通路在肠道成纤维细胞影响克罗恩病术后吻合口纤维化中的作用

项目编号： No.81500421

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 吴茸

作者单位： 东南大学

项目金额： 17万元

中文摘要： 吻合口纤维化是克罗恩病术后常见的并发症，目前还没有有效的方法预防或缓解该过程。肠道成纤维细胞在其中发挥了重要作用，但具体机制尚不明确。热休克蛋白70（HSP70）主要由成纤维细胞产生并分泌，在纤维化尤其是肠道纤维化过程中发挥着重要作用，而miR-34a则可以直接调控HSP70的表达。首先进行体外成纤维细胞培养，通过脂质体转染上调及下调miR-34a的表达，确定miR-34a/HSP70通路对小鼠成纤维细胞形态、功能及对胶原表达量等的影响；然后建立行回盲部切除、回肠升结肠吻合术IL-10基因敲除小鼠模型，通过活体转染上调及下调miR-34a的表达，确定miR-34a/HSP70通路对小鼠吻合口纤维化、胶原表达量及与纤维化相关细胞因子表达量的影响。本研究旨在阐明miR-34a/HSP70通路在肠道成纤维细胞影响克罗恩病术后吻合口纤维化中所起的作用，为克罗恩病肠纤维化的分子靶向治疗提供新的思路。

中文关键词： miR-34a/HSP70通路；克罗恩病；肠道成纤维细胞；吻合口；纤维化

英文摘要： There is no effective method to prevent or relieve the process of anastomotic fibrosis which is the most common complicantions of post-surgery Crohn’s disease patients. Fibroblast cells play an important role in this process although the mechanisms remain unknown. Heat shock protein70(HSP70), which can be directly regulated by miR-34a, is predominantly expressed in the fibroblasts and play an important part of the procession of fibrosis especially intestinal fibrosis. Firstly, the cell culture of murine fibroblast cell was established. The expressions of miR-34a were up-regulated and down-regulated by liposome transfection to determine the influence of miR-34a/HSP70 signaling pathway on morphology, function and expression of collagen in murine fibroblast cells. Secondly, the model of ileocaecal resection (ICR) in IL-10-/- mice was developed. The influence of miR-34a/HSP70 signaling pathway on anastomotic fibrosis, expression of collagen and fibrosis related cytokines were determined by transfection in vivo. The purpose of this study is to investigate the effect of miR-34a/HSP70 signaling pathway on fibrosis process of post-surgery Crohn’s disease patients so as to provide new ideas in the molecular targeted treatment of Crohn’s disease.

英文关键词： miR-34a/HSP70 pathway;Crohn's disease;intestinal fibroblast;anastomosis;fibrosis