缺血性卒中溶栓治疗的同时也会引起脑缺血再灌注损伤（CIRI），但对其机制一直未能完全阐明。我们前期通过高通量测序已经发表了大脑中动脉阻塞后再通（MCAO/R）模型大鼠皮质区的miRNA变化网络。进一步实验发现了潜在关键分子miR-758，并初步证实该分子促进细胞凋亡。同时初步确定了其可能的直接上下游调控分子：circRNA0000964和ILK。由于这三个分子在CIRI损伤中功能尚不明确，故本项目在前期工作基础上，拟使用腺相关病毒（AAV）进行体内和体外调控，揭示其生物学功能。同时通过核酸探针亲和富集和AGO2-RIP等实验确认miR-758与circRNA0000964和ILK的直接作用。最后拟使用体内营救和功能阻碍实验，验证circRNA0000964/miR-758/ILK的调控关系及在CIRI中的作用机制，并解析下游效应途径及靶点。为未来临床治疗CIRI提供新视角和可靠实验依据。

Thrombolytic therapy for ischemic stroke can also cause cerebral ischemia reperfusion injury (CIRI), but its mechanism has not been fully elucidated. We previously published the miRNA change network in the cortex of rat with middle cerebral artery occlusion recanalization (MCAO/R) by high throughput sequencing. Further experiments identified a potential key molecule, microRNA-758, and preliminarily confirmed that this molecule promotes apoptosis. Meanwhile the possible upstream and downstream regulators, circRNA0000964 and ILK, were identified. Because the functions of these three molecules in CIRI injury are still not clear, the next step in this project we will use adeno-associated virus (AAV) to regulate expressions of these three molecules respectively in vivo and in vitro, and reveal their biological functions. Then the direct interactions of miR-758 with circRNA0000964 and ILK will be confirmed by probe affinity enrichment and AGO2-RIP experiments. Finally, in vivo rescue and functional impediment experiments will be used to verify the regulatory relationships within circRNA0000964/miR-758 /ILK and its mechanism in CIRI. Furthermore the downstream effect pathways and targets will be also analyzed. Our results will possibly provide a new perspective and reliable experimental basis for the clinical treatment of CIRI.