外泌体miR-381-3p调控血管生成促进CSM神经损伤修复的作用及机制

既往研究证明外泌体miR参与血管新生及脊髓损伤后的神经修复。我们前期首先在CSM患者和动物模型减压术后发现外泌体miR-381-3p明显上调，且可促进内皮细胞增殖、迁移及血管生成。据此我们提出以下假设：外泌体miR-381-3p可调控血管生成促进CSM的神经修复。本项目拟研究CSM神经损伤修复过程中外泌体miR-381-3p的时序表达变化、表达的表观遗传学调控和血管生成相关基因相互关系；联合基因芯片和生物信息学预测分析miR-381-3p的新靶基因；GOF/LOF干预研究过表达或敲低miR-381-3p对内皮细胞增殖迁移与成管能力及相关基因表达谱变化；受精卵显微注射法构建miR-381-3p表达功能型和抑制型转基因小鼠，在体验证miR-381-3p对转基因小鼠脊髓损伤后神经修复的影响，阐明miR-381-3p调控血管形成、促进神经修复的机制，为CSM神经损伤的修复治疗提供理论依据和新靶点。

Previous studies have demonstrated that exosome miRNAs are involved in angiogenesis and nerve repair after spinal cord injury. We firstly found that exosome miR-381-3p was significantly upregulated in patients with cervical spondylotic myelopathy (CSM) and animal models after decompression. And exosome miR-381-3p could promote the proliferation, migration and angiogenesis of endothelial cells. Accordingly, we proposed the following hypothesis: abnormal expression of exosome miR-381-3p may play an important role in nerve repair of CSM via regulation of angiogenesis. This project aims to investigate the temporal changes of exosomes miR-381-3p expression during nerve injury repair in CSM, the epigenetic regulation of expression and the relationship between angiogenesis related genes. The new target genes of miR-381-3p will be predicted and confirmed by Oligo microarray and Target Scan bioinformatics, and target genes' functions will be preliminarily analyzed. The effects of overexpression or knockdown of miR-381-3p on endothelial cell proliferation, catheterization and migration, as well as related gene expression profiles will be investigated via GOF (gain-of-function) and LOF (loss-of-function) experiments. Transgenic mice with functional and inhibitory expressions of miR-381-3p will be constructed by microinjection of fertilized ovum. The effect of miR-381-3p on nerve repair of transgenic mice after spinal cord injury will be experimented. The current work is likely to lead to provide theoretical basis and new targets for the repair and treatment of nerve injury in CSM.