m6A去甲基转移酶FTO调控Seipin介导的线粒体自噬在脑缺血再灌注损伤中的机制研究

线粒体损伤是缺血性脑卒中溶栓治疗后再灌注损伤中导致神经元死亡的重要因素。FTO（Fat mass and obesity-associated protein）是RNA去甲基化酶，可以通过调控靶基因mRNA的6-甲基腺嘌呤（m6A）水平来影响靶基因mRNA的表达水平。内质网蛋白Seipin介导的线粒体自噬能够清除受损的线粒体发挥神经保护功能，但其调控机制不清。本项目前期研究发现Seipin是FTO的靶基因，FTO在脑卒中水平降低使得Seipin mRNA的m6A修饰上升而蛋白水平下降，从而抑制了Seipin介导的线粒体自噬，最终加剧了神经元的凋亡。本课题拟制备FTO和Seipin敲除、脑缺血再灌注的动物和细胞模型，应用siRNA，基因转染技术，从整体-细胞层次证明我们的假说。

Mitochondrial injury is an important factor leading to neuronal death in reperfusion injury after thrombolytic therapy in ischemic stroke. FTO （Fat mass and obesity-associated protein）is an RNA demethylase, which can regulate the expression level of target gene mRNA by regulating the level of m6A. Seipin protein is a transmembrane protein located in endoplasmic reticulum. Sepin-mediated mitochondrial autophagy.can clear damaged mitochondria and play a neuroprotective role, but its mechanism is unclear. Our preliminary data showed that Seipin was the target gene of FTO. The decrease of FTO in the level of stroke makes the m6A modification of Seipin mRNA increase and the protein level decrease, thus inhibiting the mitochondrial autophagy mediated by Seipin, and finally aggravating the apoptosis of neurons. In this project, we prepare animal and cell models of FTO and Seipin knockout, cerebral ischemia-reperfusion, and apply siRNA and gene transfection technology to prove our hypothesis from the whole cell level.