基于脂肪细胞外泌体miRNA-126激活Wnt/β-catenin通路介导电针的脑缺血耐受机制

课题组前期发现电针激活Wnt/β-catenin通路介导脑缺血耐受效应（CIT），进一步观察到电针上调血浆及脑组织miRNA-126表达、电针刺激脂肪细胞外泌体增加、脂肪细胞外泌体示踪于海马神经元、过表达miRNA-126改善神经功能。课题组提出假说：电针刺激脂肪细胞外泌体，上调囊泡内miRNA-126，后者经外泌体递送至海马神经元，下调GSK-3βmRNA，β-catenin降解减少，集聚入核介导CIT效应。拟解决：1）阐明脂肪细胞外泌体是电针诱导CIT效应的关键。2）论证miRNA-126是外泌体激活Wnt/β-catenin通路的关键。3）明确GSK-3βmRNA是miRNA-126的靶基因。从整体、组织、细胞、分子、基因水平，首次论证电针刺激脂肪细胞外泌体、上调miRNA-126、激活Wnt/β-catenin通路介导CIT效应的机制，为缺血性脑卒中的中医脑保护研究提供科学依据。

Our group found previously that electro-acupuncture activated Wnt/beta-catenin pathway mediated cerebral ischemic tolerance effect (CIT), and we intend to further elucidate its mechanism. Preliminary experiments showed that electro-acupuncture up-regulated the expression of microRNA-126 in plasma and brain tissue, electro-acupuncture stimulated the increase of adipocyte exosomes, adipocyte exosomes can be traced to hippocampal neurons, and over-expression of microRNA-126 improved nerve function. Our hypothesis is that electroacupuncture stimulates the secretion of adipocytes, promotes the expression of microRNA126 in vesicles, and microRNA126 down-regulates GSK-3 beta gene, reduces the degradation of beta-catenin and mediates CIT effect in nuclei. To solve: 1) To clarify that adipocyte exosome is the key to CIT induced by electroacupuncture. 2) To demonstrate that microRNA-126 is the key to the activation of Wnt/beta-catenin pathway by exosomes. 3) To clear that GSK-3beta is the target gene of microRNA-126. The mechanism of electroacupuncture stimulating adipocyte exosome, up-regulation of microRNA-126, activation of Wnt/beta-catenin pathway mediating CIT effect is demonstrated for the first time at the level of whole, tissue, cell, molecule and gene, which provides a scientific basis for the traditional Chinese medicine study about ischemic stroke.