MicroRNA-128b-3p靶向wnt3a/β-catenin信号通路调控糖尿病小鼠角膜上皮及神经损伤修复的作用与机制研究

Diabetic keratopathy is one of the common ocular complications of diabetes mellitus, and its clinical interventions are limited. However, the current mechanism and theoretical study of diabetic keratopathy are still inadequate. Our previous studies showed that the wnt3a in the corneal epithelium was decreased in diabetic mice. After screening the possible microRNAs, we found that the expression of microRNA-128b-3p in corneal epithelium of diabetic mice increased significantly. The exogenous microRNA-128b-3p antagomir could significantly improve the repair rate of corneal epithelium and increase the sensitivity of corneal nerves in diabetic mice. Therefore, we put forward the hypothesis that the microRNA-128b-3p may be a potential factor for the diabetic keratopathy: It may regulate diabetic keratopathy by targeting Wnt3a signaling pathway. In this project, the models of corneal epithelium and nerve injury in type 2 diabetic mice are used. The mechanism of the microRNA-128b-3p/wnt3a for corneal nerve and corneal epithelium repairing in diabetes is to be further investigated. The conclusion of this study will help clarify the mechanism of microRNA-128b-3p regulates the repair of corneal epithelium and nerve injury by targeting Wnt3a/β-catenin signaling pathway in diabetic mice, and provide a new intervention strategy for the treatment of diabetic keratopathy.

糖尿病性角膜病变是糖尿病常见眼部并发症之一，临床干预手段十分局限。目前糖尿病性角膜病变的机制及理论研究相对不足。申请者前期研究发现：糖尿病小鼠角膜上皮wnt3a水平降低，对其可能的microRNA筛选后发现microRNA-128b-3p在糖尿病小鼠角膜上皮表达水平明显增高。外源性抑制microRNA-128b-3p可显著促进糖尿病小鼠角膜上皮及神经损伤修复。因此，从前期预实验结果可以推测microRNA-128b-3p可能通过靶向wnt3a信号通路对糖尿病角膜神经上皮损伤修复进行调控。本项目拟采用2型糖尿病小鼠角膜神经上皮损伤模型，深入探讨microRNA-128b-3p/wnt3a对糖尿病角膜神经上皮损伤修复的作用及机制。本研究结论将有助阐明microRNA-128b-3p靶向wnt3a/β-catenin信号在调控角膜缘干细胞更新、角膜上皮以及神经损伤修复过程中的作用机制，为糖尿病性角膜病变的治疗提供新的干预策略。

糖尿病性角膜病变是糖尿病常见眼部并发症之一，临床干预手段十分局限。目前糖尿病性角膜病变的机制及理论研究相对不足。在本研究中，我们利用糖尿病小鼠模型、TKE2细胞和体外培养的三叉神经节细胞，使用免疫荧光、WB、RT-PCR等手段摸索相关通路Wnt信号通路以及miRNA-128b-3p等在糖尿病小鼠角膜的表达。并通过结合Insulin、ADPN等因子进一步扩展研究wnt信号通路在糖尿病小鼠角膜损伤修复的作用和机制。结果证实wnt通路在糖尿病小鼠角膜神经损伤过程中发挥了重要作用，初步阐明wnt信号通路和小分子在糖尿病角膜损伤修复的机制。根据筛选的wnt信号通路，筛选几种相关因子，进行进一步验证探究wnt的激活或作用靶点，通过Insulin和ADPN等因子相结合，寻求可行的，具有转化意义的wnt信号通路相关的活性因子，以期对解决临床问题和后续的基础实验研究提供思路和契机。本研究结论将有助阐明小分子和wnt信号通路在调控角膜缘干细胞更新、角膜上皮以及神经损伤修复过程中的作用机制，为糖尿病性角膜病变的治疗提供新的干预策略。