基于miRNA-155/YY1/p53基因网络模型研究电针预处理降低缺血性卒中脑微血管内皮细胞凋亡的分子机制

电针预处理可以诱导脑缺血耐受，但其机制尚不明确。我们前期利用生物信息技术构建缺血性脑卒中相关miRNA/TF靶基因调控网络，建立细胞模型，发现miRNA-155在缺血性脑卒中的发展过程中发挥“开关”作用并靶向YY1/p53通路促进脑微血管内皮细胞凋亡；电针预处理可调控p53通路降低脑缺血再灌注损伤区域细胞凋亡。我们推测：电针预处理可以抑制miRNA-155/YY1/p53通路，减轻大鼠脑微血管内皮细胞凋亡，诱导脑缺血耐受。本课题分别从动物水平利用行为学、基因扩增及分子印迹技术，证实电针预处理通过下调miRNA-155靶向YY1/p53负反馈通路调节I/R大鼠脑微血管内皮细胞凋亡；从细胞水平利用慢病毒转染、报告基因及免疫共沉淀技术，验证miRNA-155/YY1/p53通路在脑微血管内皮细胞凋亡中的具体调节机制。旨在为缺血性脑卒中的发病机理提供新的理论依据，为临床精准预防和治疗提供新思路。

Electroacupuncture pretreatment can induce cerebral ischemic tolerance, but its mechanism is not yet clear. We have previously studies ignored the overall changes in genomics from a single molecular perspective. The applicant has previously constructed a miRNA-mediated feedback and feedforward circulation network related to ischemic stroke and found that miR-155 plays a ."switch" role in the development of the disease ,which can target YY1/p53 to promote cerebral microvascular endothelial cell apoptosis. We also found that electroacupuncture preconditioning can regulate the p53 pathway to reduce cerebral ischemia reperfusion injury. We speculate that electroacupuncture pretreatment can inhibit the miRNA-155/YY1/p53 signaling pathway to reduce cerebral microvascular endothelial cell apoptosis, then induce cerebral ischemic tolerance. Based on this, we will use the techniques of gene amplification and molecular imprinting at the animal level to verify that electroacupuncture pretreatment down-regulats miR-155 targeting the YY1/p53 negative feedback pathway to regulate the apoptosis of cerebral microvascular endothelial cells in I/R rats. And then, We will use lentiviral transfection, reporter gene and mRNA-IP techniques to verify the specific regulation mechanism of miR-155/YY1/p53 pathway in the cerebral microvascular endothelial cell apoptosis. This research aims to provide a new theoretical basis for the pathogenesis of ischemic stroke, and provide new ideas for clinical precise prevention and treatment.