神经保护剂T-006调控线粒体促进神经元再生作用机理研究

Neurodegenerative diseases are a group of diseases that are characterized by extensive neuronal loss and progressive degeneration of structure and function as major pathological features, which causes a great mental and medical burden to the upcoming ageing society. If we can effectively promote the regeneration of neurons, restore the number and function of damaged neurons, it is expected to fundamentally reverse the disease course, but few drugs were used in clinic. Promoting mitochondrial biogenesis and dynamics can improve mitochondrial function, which is an effective strategy for stimulating neuronal regeneration. The neuroprotective agent T-006, which was designed and synthesized by the applicant, can induce the neurogenesis in Parkinson’s disease (PD) and transient middle cerebral artery occlusion (t-MCAO) rats and promote mitochondrial biogenesis, and show potential therapeutic effects in PD and stroke as well as the other neurodegenerative disorders. However, how T-006 exerts its role in promoting nerve regeneration and its exact mechanism has not been systematically revealed. Thus, this project will deep investigate the effects of overexpression and silencing of PGC1α on mitochondrial biosynthesis, division/fusion and nerve regeneration of T-006. Moreover, To elucidate the role of T-006 in improving mitochondrial function and promoting neuronal regeneration by using the quantitative proteomics technology with iTRAQ labeling in combination with 2D-LC-MS/MS method. We hope to promote the compound T-006 into clinical candidate drug and provide a theoretical basis for the development of new neurogenesis drugs through the above theoretical and experimental researches.

神经退行性疾病是一类以广泛的神经元丢失或结构和功能性进行性变性为主要病理特征的疾病总称，给即将到来的老龄化社会造成极大的精神和医疗负担。促进神经元再生、恢复受损神经元数量和功能有望从根本上逆转疾病病程，但目前此类临床药物极其缺乏。促进线粒体生物合成及动力学平衡改善线粒体功能是一种重要的促神经元再生策略。申请人前期设计合成的神经保护剂T-006可促进帕金森病(PD)和脑中风(t-MCAO)大鼠脑区神经元再生，并促进线粒体生物合成，展现出独特的抗神经退行性疾病治疗潜力。但T-006是如何发挥促神经再生作用以及其具体的作用机理是什么还不明确。本项目将通过体内外研究PGC1α过表达和沉默后对T-006促线粒体生物合成、分裂/融合及神经再生的影响；并借助iTRAQ标记联合2D-LC-MS/MS定量蛋白质组学技术，阐明T-006改善线粒体功能促进神经元再生机制，为开发新型促神经再生药物提供理论基础。

促进神经元再生、恢复受损神经元数量和功能有望从根本上逆转疾病病程，但目前此类临床药物极其缺乏。促进线粒体生物合成及动力学平衡改善线粒体功能是一种重要的促神经元再生策略。本项目结合体内外研究手段发现T-006显著促进6-OHDA大鼠相关脑区DA、DCX及Nestin标记的神经元再生；并且T-006还能够上调6-OHDA大鼠脑内PGC1α的表达；进一步机理研究发现T-006能够促进NSCs干细胞线粒体的生物合成及调节其分裂/融合水平从而改善线粒体功能，表明T-006可以通过激活PGC1α改善线粒体功能发挥其促神经再生作用。此外，神经再生受到多种调控因子的影响，其中肿瘤坏死因子(TNF-α)等炎症因子被认为是影响神经再生的一种重要因素。研究表明TNF-α能够负性调控成年小鼠脑内海马神经祖细胞的增殖，而TNF-α等炎症因子的减少则能刺激神经元的再生。基于上述研究背景，本项目进一步评估了T-006抗神经炎活性，发现T-006能够降低LPS引起的细胞死亡及炎症因子的产生，并且进一步机理研究发现T-006抗神经炎活性与其抑制TLR4介导的MyD88/NF-κB信号通路有关。