采用ADSCs诱导分化的SCLCs复合去细胞神经修复周围神经损伤是目前国内外研究的热点。申请人前期研究发现ADSCs诱导分化率较低，且SCLCs促神经再生的能力明显不如SCs。针对此难题，申请人前期应用miRNA芯片筛选验证了分化前后miRNA的差异表达谱，靶基因分析发现该分化过程和细胞自噬相关。在预实验中，我们发现自噬激活具有促进ADSCs分化为SCLCs的作用。进一步RNA表达谱分析结果显示自噬可能通过调控Notch信号通路参与此过程，我们初步发现Atg5依赖的自噬可降低Notch信号通路抑制蛋白Numb表达。在此基础上，本项目将深入研究自噬是否提高SCLCs促进背根神经元轴突生长及在大鼠体内复合去细胞神经促进坐骨神经缺损修复，进一步探讨自噬对ADSCs诱导分化为SCLCs的调控作用，并通过免疫蛋白共沉淀法、免疫荧光法等证明自噬通过降解Numb蛋白激活Notch信号通路

The research of using acellular nerve graft laden with SCLCs differentiated from ADSCs to repair peripheral nerve injury is always the hot issue in the international research field. In the preliminary research, the applicant found that the number of SCLCs differentiated from ADSCs was insufficient to meet the demands of replantation, and the ability of SCLCs to promote nerve regeneration was significantly inferior to SCs. In accordance with this problem, the applicant profiled miRNA expression before and after SC-like induction using miRNA microarray essay and verified the results using qRT-PCR. Moreover, KEGG analysis showed that autophagy might involve in the process of ADSCs differentiating into SCLCs. In preliminary experiments we found autophagy activation could promote the process of ADSCs differentiating into SCLCs. Spectrum analysis further found that in high expression of Atg5 cells, notch signaling pathway is activated most significantly, and we found Atg5 could suppress the expression of numb, an inhibitor of notch signaling pathway. On this basis, we will examine whether autophagy could promote the axon growth of DRG, and sciatic nerve regeneration. Also, using immunofluorescence, immune protein co-precipitation and other methods, we try to determine whether autophagy could activate notch signaling pathway through degrading numb.