lncRNA-BIRF/miR-330-5p/GLT-1轴在脑缺血预处理诱导脑缺血耐受中的关键作用及机制研究

研究发现脑缺血预处理(CIPC)可通过上调GLT-1诱导脑缺血耐受，但其上游调制机制尚未阐明。本课题预实验结果显示，lncRNA-BIRF、miR-330-5p分别参与氧-葡萄糖剥夺(OGD)预处理上调GLT-1的调控，通过测序和生物信息学分析提示三者之间存在密切相关性。据此推测“CIPC可能通过lncRNA-BIRF/miR-330-5p/GLT-1轴诱导脑缺血耐受”。针对该设想，本项目拟应用报告基因分析、高效液相色谱等多种技术，研究lncRNA-BIRF在CIPC上调GLT-1诱导脑缺血耐受中的关键作用，论证miR-330-5p靶向调控GLT-1参与CIPC诱导脑缺血耐受，阐明CIPC通过lncRNA-BIRF竞争性结合miR-330-5p，靶向调控GLT-1，诱导脑缺血耐受的分子机制。研究结果将丰富对脑缺血耐受诱导机制的认识，为脑缺血性疾病防治研究提供新思路和新的候选靶标。

Cerebral ischemic preconditioning (CIPC) can induce brain ischemic tolerance by up-regulating GLT-1 expression. However, the upstream regulation mechanisms in this process are still unclear. Our preliminary experiments results suggest that lncRNA-BIRF and miR-330-5p both contribute to the GLT-1 up-regulation induced by OGD preconditioning. RNA-seq and bioinformatics analysis show that there is a close relationship among lncRNA-BIRF, miR-330-5p and GLT-1. Taken together, we hypothesize that lncRNA-BIRF/miR-330-5p/GLT-1 axis may participate in the induction of brain ischemic tolerance induced by CIPC. The present project is aimed to indentify the hypothesis by using double luciferase reporter gene analysis, high performance liquid chromatography and other methods. At first, the study will elucidate the key role of lncRNA-BIRF in the induction of brain ischemic tolerance by CIPC via GLT-1 up-regulation. Then demenstrate that miR-330-5p participates in the induction of brain ischemic tolerance by CIPC through targetting GLT-1. In the end, we will clarify the underlying mechanism of CIPC inducing brain ischemic intolerance through lncRNA-BIRF competitively binding to miR-330-5p to up-regulate GLT-1 expression. The results of the project will enrich the understanding of the induction mechanism of brain ischemia tolerance, and present new ideas and candidate target for studying the precaution and treatment of brain ischemia.