RhoGDIα对小鼠脑缺血再灌注损伤的神经保护功能及其机制研究

Cerebral infarction is a serious threat to human health disease, thrombolysis treatment to restore ischemic brain tissue perfusion to save the dying neurons, but also cause reperfusion injury resulted in its death. Therefore, the study of the nerve repair function after cerebral ischemia-reperfusion injury and its mechanism has important significance. Our previous study found that the eIF5A1 through RhoGDI alpha promote the spinal neuronal survival and neurite outgrowth, neuroprotective and neural plasticity, and prove that promotes cell survival and expression in spinal cord regeneration content projection degree RhoGDIα positive correlation. So the speculation RhoGDI alpha has a tremendous potential as therapeutic targets. And the molecular function after cerebral ischemia reperfusion injury and the mechanism is unreported. This project intends to establish a suture method in mice cerebral ischemia and reperfusion model, in different brain regions as well as the expression of specific cell types (such as neurons, astrocytes, etc.) RhoGDI alpha analyzed. While using the human body and have therapeutic potential for toxic and side effects AAV viral vector. RhoGDI alpha effects of specific brain regions and cell types in vivo, study on brain cells survive, damage repair and the influence of neural plasticity. The possible simultaneous detection of the promoter region of DNA methylation changes, to find its upstream translation regulation mechanism, with good clinical and social significance.

脑梗死是一种严重威胁人类健康的疾患，溶栓治疗使缺血脑组织恢复灌注以挽救濒临死亡的神经细胞，同时也造成再灌注损伤而导致死亡。因此，研究脑缺血再灌注损伤后的神经修复及其机制具有重要的意义。我们前期的研究结果发现，eIF5A1通过RhoGDIα促进脊髓神经元存活及突起生长，具有神经保护功能，并证明其促进神经细胞存活和突起再生的程度与RhoGDIα的表达含量正相关。且该分子在脑缺血再灌注损伤后的功能及机制尚无报道。本项目拟建立线栓法小鼠脑缺血再灌注模型，对不同脑区中特定细胞类型的RhoGDIα表达进行分析。同时运用具有治疗潜能且对人体毒副作用小的AAV病毒载体。针对RhoGDIα影响的具体脑区和细胞类型进行调控，研究其对脑细胞存活、损伤修复和神经可塑性的影响。同时检测其可能的启动子区DNA甲基化变化，探寻其上游调控机制，为可能的后续临床应用提供支持。

环状RNA（circRNAs）是由头尾剪接产生的，在所有多细胞生物中广泛表达。它们的重要生物学功能越来越受到人们的重视。磷脂酰肌醇-3,4,5-三磷酸[PI（3,4,5）P3]是磷脂酰肌醇4-磷酸[PI（4）P]和磷脂酰肌醇4,5-二磷酸[PI（4,5）P2]的磷酸化衍生物，它们在质膜（PM）中组建并激活AKT以促进细胞存活。ORP5锚定在内质网（ER-PM）的接触位点，充当PI（4）P和PI（4,5）P2/磷脂酰丝氨酸（PS）的交换剂。在这里，对感觉运动皮层的脂质组学分析显示短暂性大脑中动脉阻塞（tMCAO）扰乱了磷脂酰肌醇（PIs）和PS在PM和ER之间的稳态。条件敲除小鼠显示ORP5参与了这种异常分布。敲除ORP5基因可显著抑制细胞凋亡和自噬。RNA测序和RNA下拉分析证实了一种竞争性内源性RNA（ceRNA）途径，其中circ_0001449与miR-124-3p和miR-32-5p促进Osbpl5翻译。我们的数据表明，circRNA_0001449通过ORP5调节膜内稳态，并参与AKT的生存途径。