基于外泌体介导神经元-雪旺细胞信号通讯的糖络宁防治DPN的作用机制研究

Diabetes peripheral neuropathy (DPN) is one of the complications of diabetes.Neuron-Schwann cells functional injury is a key factor in the demyelination and axonal degeneration. Abnormal Neuron-Schwann cells signal communication mediated by Exosomes (EXOs) is associated with DPN pathogenesis. Our researches previously found that Tangluoning has definite clinical effect in DPN, which involves in the pathogenesis such as oxidative stress and endoplasmic reticulum stress. However, the role of EXOs in the pathogenesis of DPN is unclear. This project intends to conduct DPN rat model and a co-culture model of dorsal roots ganglia neuron and Schwann cells in a high glucose environment, and also methods such as proteomics, bioinformatics and microarray chip and technology as QRT - PCR, laser scanning confocal microtechnic and Western Blot were adopted. Through the above methods,the correlation between DPN and dysfunction of neuron-Schwann cells caused by abnormal intercellular signal communication mediated by EXOs was analyzed in vivo and vitro experiments. Based on this new perspective of neuron and Schwann cells signal communication mediated by EXOs, the project reveal the mechanism and target of Tangluoning in the prevention and treatment of DPN, and provides a scientific basis for the development of new drugs for DPN.

糖尿病周围神经病变（DPN）是糖尿病常见并发症之一，神经元-雪旺细胞（SCs）的功能受损是神经病变脱髓鞘和轴突退化的关键因素，外泌体（EXOs）介导的神经元-SCs信号通讯与DPN发病机制相关。本课题组前期研究发现糖络宁防治DPN临床疗效确切，涉及氧化应激和内质网应激等DPN发病机制，而EXOs如何参与DPN的发病机制尚不明确。本项目拟利用DPN大鼠模型和高糖环境背根神经节神经元（DRGn）-SCs共培养细胞模型，采用蛋白质组学、生物信息学和微阵列芯片研究方法和qRT-PCR、激光共聚焦和WB等技术手段，分别从体内、体外两个侧面和组织、细胞、蛋白和基因不同层次,阐释EXOs介导的神经元-SCs信号通讯-功能异常-DPN的作用机制，预期基于EXOs介导神经元-SCs信号通讯这一新视角，揭示糖络宁防治DPN的作用机制和靶点，为防治DPN的中药新药开发提供科学依据。

糖尿病周围神经病变（DPN）是糖尿病常见并发症之一，神经元-雪旺细胞（SCs）的功能受损是神经病变脱髓鞘和轴突退化的关键因素，外泌体（EXOs）介导的神经元-SCs信号通讯与DPN发病机制相关。本课题组前期研究发现糖络宁防治DPN临床疗效确切，涉及氧化应激和内质网应激等DPN发病机制，本研究证明EXOs装载的蛋白异常参与DPN。本项目利用DPN大鼠模型和高糖环境背根神经节神经元（DRGn）-SCs共培养细胞模型，采用蛋白质组学、生物信息学和Small RNA测序技术等研究方法，分别从体内、体外两个侧面和组织、细胞、蛋白不同层次，围绕DPN脱髓鞘和轴突萎缩，筛选筛选Laminin211、Integrin α6β1和14-3-3beta/alpha为标志蛋白，Laminin211/Integrin α6β1调控的rac1/cdc42和cofilin通路、14-3-3beta/alpha/RhoA通路为关键环节，研究DPN的发病机制及糖络宁重塑细胞外微环境，促进轴突生长和髓鞘形成的作用机制，为防治DPN的中药新药开发提供科学依据。此外，本项目为血浆外泌体作为DPN的诊断载体提供了初步科学依据。