星形胶质细胞中HSPA12B调节SSeCKS在多发性硬化炎症过程中作用机制的研究

Multiple sclerosis (MS) is a chronic and potentially highly disabling disorder with considerable social impact and economic consequences. Inflammatory activation of astrocyte mediated by SSeCKS played an important role in multiple sclerosis. Our previous studies had found that HSPA12B interacted with SSeCKS involved in astrocyte activation and migration induced by inflammation. In this study, EAE, the common animal model of MS was used to study the expression and interaction between SSeCKS and HSPA12B during inflammation and astrocyte activation in MS. To clarify the binding sites of SSeCKS and HSPA12B which regulated the activity of SSeCKS and the inflammatory activation of astrocyte , the inflammatory activation model of astrocyte was established. By co-culturing of astrocytes and oligodendrocytes to demonstrate the impact of the interaction of SSeCKS and HSPA12B to the molecul related to myelination. Finally, to clarify the molecular mechanisms of demyelinating process in MS, and to provide a new way for the prevention and treatment of MS.

多发性硬化（multiple sclerosis，MS）是一种慢性疾病，具有高潜在致残率，伴随着相当大的社会影响和经济后果。SSeCKS介导星形胶质细胞炎性激活在MS过程中发挥重要作用。我们先前的研究发现：与SSeCKS相互作用的HSPA12B参与炎症诱导的星形胶质细胞的激活和迁移过程。本课题拟通过MS动物模型EAE，研究SSeCKS和HSPA12B在MS炎症和星形胶质细胞活化过程中的表达及相互作用；通过研究星形胶质细胞炎性活化模型，阐明HSPA12B通过与SSeCKS不同位点的相互结合，调节SSeCKS的活性，从而调控星形胶质细胞炎性激活；通过星形胶质细胞与少突胶质细胞共培养，阐明两者相互作用对髓鞘形成相关蛋白表达的影响，从而探讨两者在MS脱髓鞘过程中的作用和分子机制，籍以为MS的预防和治疗提供新的方法。

目的：Src抑制的蛋白激酶C底物（SSeCKS）参与多发性硬化动物模型EAE的炎性星形胶质细胞诱导的少突胶质细胞凋亡。最近通过酵母双杂交发现了SSeCKS 与HSPA12B存在相互作用。我们的研究旨在阐明SSeCKS和HSPA12B在EAE星形胶质细胞激活中的相互作用及其潜在的机制。.方法：运用免疫印迹和免疫荧光分析HSPA12B和SSeCKS在EAE中的表达和定位，以及免疫共沉淀分析两者在EAE星形胶质细胞激活中的相互作用。运用ELISA检测致炎细胞因子TNF-α 和 IL-1β的释放。构建HSPA12B和SSeCKS过表达和抑制作用的载体检测MAPK信号通路对致炎细胞因子产生过程的作用。分别使用JNK和P38的抑制剂SP600125 和SB202190分析其在星形胶质细胞激活中的作用。构建全长和片段SSeCKS检测其与HSPA12B的结合位点。.结果：通过检测我们发现在EAE星形胶质细胞活化过程中SSeCKS和HSPA12B存在相互作用，HSPA12B可能通过调节SSeCKS的表达和活化从而调控LPS刺激的星形胶质细胞激活和炎症因子TNF-α 和 IL-1β的释放。HSPA12B可能通过介导JNK 和p38信号通路, 而不是通过介导ERK1/2 信号通路调节星形胶质细胞的激活。并且HSPA12B通过N端1-330和C端1278-1596位点与SSeCKS结合。.结论：HSPA12B通过调节SSeCKS的表达和活化从而调控LPS刺激的星形胶质细胞激活和炎症因子TNF-α 和 IL-1β的释放。HSPA12B通过介导JNK 和p38信号通路, 而不是通过介导ERK1/2 信号通路调节星形胶质细胞的激活。并且HSPA12B通过N端1-330和C端1278-1596位点与SSeCKS结合。EAE过程中HSPA12B与SSeCKS相互作用的调节机制可能有望为MS提供新的治疗方法。