LncRNAK02111沉默Wnt5a促进糖尿病皮肤成纤维细胞凋亡的作用及机制

项目名称： LncRNAK02111沉默Wnt5a促进糖尿病皮肤成纤维细胞凋亡的作用及机制

项目编号： No.81471034

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 严励

作者单位： 中山大学

项目金额： 75万元

中文摘要： 糖尿病常伴有皮肤损害和修复障碍，皮肤成纤维细胞凋亡在其中发挥关键作用。课题组前期研究发现，lncRNAK02111在糖尿病皮肤组织异常高表达，且与成纤维细胞凋亡密切相关；但lncRNAK02111如何介导皮肤成纤维细胞凋亡，目前尚不明确。我们进一步通过RNA-pull down实验发现lncRNAK02111与抗凋亡蛋白Wnt5a相结合。本项目拟在前期研究基础上，采用体内实验和体外研究结合的方法，以lncRNAK02111为切入点，从细胞、组织和整体三个层次深入研究。采用RNA-IP、Deletion mapping等手段首先证实lncRNAK02111直接结合Wnt5a，继而探讨lncRNAK02111功能性沉默Wnt5a、抑制下游PKA-CREB 抗凋亡途径、促进成纤维细胞凋亡的作用及机制。本研究有望揭示糖尿病皮肤病变发生的表观遗传调控机制，为糖尿病皮肤病变治疗确立新靶点提供科学依据。

中文关键词： 糖尿病足；成纤维细胞；凋亡；长链非编码RNA；Wnt5a

英文摘要： Diabetes is associated with impaired wound healing,and fibroblast cell apoptosis of skin tissues plays a key role in the process. In our previous study, we applicated LncRNAs microarray technology to detect and screen the specific lncRNAs of diabetic skin tissues, and validated them using quantitative real-time polymerase chain reaction.Results demonstrated that lncRNAK02111 was significantly upregulated in skin tissues of both diabetic patients and rats,and was closely associated with fibroblast cell apoptosis.Further RNA pull-down experiments showed that lncRNAK02111 combined with the anti-apoptotic protein Wnt5a. Therefore, we speculate that lncRNAK02111 combines with Wnt5a directly and silences its function,then inhibits its anti-apoptotic function through PKA-CREB pathway in diabetic fibroblast cells. All will finally lead to skin damage and impaired wound healing.The aim of the present study was to firstly confirm the direct combination effect of lncRNA K02111 with the protein Wnt5a, and then explore the involved PKA-CREB signaling pathway of Wnt5a function silence in the process of fibroblst apoptosis. This research may help to discover the new mechanism of fibroblast cell apoptosis in diabetic skin, and represent potential targets for drug interventions to improve diabetic wound healing.

英文关键词： diabetic foot;fibroblast;apoptosis;lncRNA;Wnt5a