TIMP1对SOD1突变导致ALS血脊屏障损伤的调节作用及机制研究

肌萎缩侧索硬化症（ALS）是以运动神经元进行性缺失为主要病理特征的致死性神经变性病，目前尚无有效治疗手段。近年来发现血脊屏障功能异常早于ALS神经系统病变，提示早期靶向血脊屏障可作为ALS潜在治疗策略。申请者前期工作发现TIMP1可通过减轻血脑屏障损伤在创伤性脑外伤中发挥保护作用；采用ALS患者及动物样本，结合GEO数据库分析结果提示TIMP1可能参与ALS进展；给予TIMP1重组蛋白可逆转不同生物特征SOD1突变体介导的血管内皮细胞屏障功能损伤。在此基础上，本项目拟采用SOD1G93A小鼠、原代及血管内皮细胞系，①明确TIMP1对ALS血脊屏障功能失调及神经元病理损伤的改善作用；②阐明TIMP1对血脊屏障调控的分子机制，重点关注细胞因子调控的RhoA抑制作用和基质金属蛋白酶抑制活性。本项目旨在探索以TIMP1进行早期干预改善ALS患者治疗效果的可能性，为ALS药物研发提供新思路。

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease with progressive loss of motor neurons as the major pathological feature, and currently there is no effective treatment. In recent years, it has been found that the blood-spinal cord barrier function is compromised prior to neuropathy in ALS, suggesting that early targeting the blood-spinal cord barrier can be a potential therapeutic strategy for ALS. Our previous work has shown that TIMP1 displays a protective role in traumatic brain injury by ameliorating blood-brain barrier disruption. The analysis results of ALS patients and animal samples, and GEO database suggest that TIMP1 may be involved in the progression of ALS. Administration of recombinant TIMP1 could reverse the vascular endothelial barrier function impairment induced by SOD1 mutants with different biochemical characteristics. On this basis, this project intends to use SOD1G93A mouse, primary and microvascular endothelial cell lines ① to clarify the effect of TIMP1 on blood-spinal cord barrier dysfunction and neuronal pathological damage in ALS; ② to investigate the molecular mechanism underlying TIMP1 regulation on the blood-spinal cord barrier, especially focusing on the RhoA GTPase inhibitory effect and matrix metalloproteinase inhibitory activity. This project aims to explore the possibility of early intervention with TIMP1 to improve the treatment effect of ALS patients, and to provide novel ideas for the development of ALS drugs.