益肾活血法中药调控星形胶质细胞和补体系统治疗视神经脊髓炎的神经保护作用机制研究

Neuromyelitis optica(NMO) is autoimmune disease. The incidence of NMO was higher.NMO has higher relapse rate,and cause severe disability. Some of NMO severe neurological damage is irreversible in the first attack. On the basis of the overview to pathogenesy theory of Traditional Chinese Medicine (TCM) for NMO, we set up the method of reinforcing kidney and activating blood therapy.From the aspects of pathogenesis and pathological changes localization, we interpreted that the basic pathogenesis characteristic was brain marrow and collateral injury due to deficiency of kidney-essence, function disharmony of entrails, recurrence of exogenous evil. On the other hand, the direction to treatment according to reinforcing kidney and activating blood was brought forward. In the early research, we found Chinese medicine has antihypoxia and neuroprotective effect. Part of the results published in Journal of Neurochemistry. On the basis of previous research results, we carry out the following research. Rats intracerebral injection NMO-IgG will use to establish model of NMO,and the NMO cell model will establish with NMO-IgG co-cultured with astrocytes in the system. The astrocytes was key target in the pathogenesis of NMO.We research the roles of astrocyte activation immune reaction in NMO nerve injury,and astrocytes induced neurotoxicity of complement.We will research the mechanism of astrocytes in NMO neurotoxicity in animal, tissue, cell, molecular. And explore the neuroprotective mechanism of NMO against immune injury about Yishen Huoxue Prescription. From the analysis of the neurotoxicity mechanism and neuroprotective effect in NMO with TCM prescription, we will supply the basis for seaking the preponderance in treating NMO with the TCM and exploring the best opportunity of clinical intervention with the TCM.

视神经脊髓炎(NMO)是中枢神经系统自身免疫性疾病。我国NMO发病率高、复发率高、致残程度重、疗效较差。本研究提出NMO肾精不足、浊毒瘀痹经脉的病机理论，创立益肾活血的治法，采用具有益肾活血作用的中药复方益肾达络饮治疗NMO，研究其作用机制。课题组前期有关益肾活血中药干预实验性自身免疫脑脊髓炎(EAE)缺氧和免疫紊乱的神经保护作用的研究，取得可喜的结果，部分成果发表在J Neurochem杂志(影响因子4.281)。本研究在前期研究成果基础上，采用NMO-IgG作用于EAE的方法制备NMO动物模型和细胞模型，以NMO发病的关键靶点---星形胶质细胞为切入点，从动物、组织、细胞、分子多个维度研究NMO星形胶质细胞介导的免疫反应和激活补体导致的神经毒性作用的机理，揭示NMO的发病机制及其益肾活血方药的神经保护作用。深入探求中医药治疗NMO的作用机理，为中医药治疗NMO提供理论依据。

视神经脊髓炎谱系疾病(NMOSD；既往称为视神经脊髓炎，即NMO，2015年国际NMO诊断小组取消了NMO的单独定义，将NMO整合入更广义的NMOSD疾病范畴中。自此，统一命名为NMOSD，我国自2016年统一使用NMOSD的病名)是中枢神经系统自身免疫性疾病。我国NMOSD发病率高、复发率高、致残程度重、疗效较差。本研究提出NMOSD肾精不足、浊毒瘀痹经脉的病机理论，创立益肾活血的治法，采用具有益肾活血作用的中药复方益肾达络饮治疗NMOSD，研究其作用机制。. 我们建立了NMOSD动物模型----实验性自身免疫性脑脊髓炎模型(EAE)，通过对星形胶质细胞介导的NMOSD免疫损伤作用与脱髓鞘改变、神经细胞损伤的研究，明确了星形胶质细胞在免疫反应中的关键作用；通过对补体通路参与的神经毒性作用机制研究，发现在众多补体因子中CD59和factor B在NMOSD发病过程中密切相关，星形胶质细胞介导了补体系统在NMOSD的神经损伤；星形胶质细胞可介导多种炎性因子异常表达，包括NF-κB、P65、Th17、Hsp70等，其中Th17细胞是最重要的炎性细胞之一，Hsp70和miR-374c可以调控Fas的表达，下调miR-374c和Hsp70可通过诱导EAE中Fas的表达促进Th17细胞分化；星形胶质细胞还介导了大量氧化应激因子高表达，包括CytoC、NOX2等。这些炎症因子和氧化应激因子高表达，继发线粒体凋亡相关蛋白表达可能是NMOSD脱髓鞘后不可逆神经损伤的深层次原因；项目研究还发现，EAE神经损伤修复过程中，神经干细胞起着重要的作用。神经干细胞的增殖和分化受其内在基因的调控，Mash1被认为是正性分化调节基因，Hes1被认为是负性分化调节基因，两者之间可发生拮抗作用。. 具有益肾活血作用的中药复方益肾达络饮通过调节NMOSD模型星形胶质细胞、炎症因子、补体，修复混乱的免疫功能、保护髓鞘；通过调节Mash1和Hes1基因表达，促进神经干细胞分化，达到修复神经功能的作用。本项目的研究为进一步的NMOSD治疗研究确定了治疗靶点，中药防治NMOSD的临床应用奠定了良好的基础。. 通过项目研究，已发表跟本项目相关学术论文6篇，其中SCI源期刊论文2篇(其中1篇发表在Int J Biol Macromol杂志IF 5.162)，中文核心收录4篇。