METHODS: The interaction between microglia and AD was investigated in vitro and in vivo. RNA-binding protein immunoprecipitation (RIP) was used to investigate the mechanisms of miR-223 and YB-1. The association between microglia derived exosomal YB-1/miR-223 axis and nerve cell damage were assessed using Western blot, immunofluorescence, RT-PCR, ELISA and wound healing assay.
RESULTS: Here, we reported AD model was responsible for the M1-like (pro-inflammatory) polarization of microglia which in turn induced nerve cell damage. While M2-like (anti-inflammatory) microglia could release miR-223-enriched EXO which reduced neuroinflammation and ameliorated nerve damage in AD model in vivo and in vitro. Moreover, YB-1 directly interacted with miR-223 both in cell and EXO, and participated in microglia exosomal miR-223 loading.
CONCLUSIONS: These results indicate that anti-inflammatory microglia-mediated neuroprotection form inflammatory damage involves exporting miR-223 via EXO sorted by YB-1. Consequently, YB-1-mediated microglia exosomal sorting of miR-223 improved the nerve cell damage repair, representing a promising therapeutic target for AD.
方法:在体外和体内研究了小胶质细胞与AD之间的相互作用。RNA结合蛋白免疫沉淀(RIP)用于研究miR-223和YB-1的机制。使用Westernblot评估小胶质细胞来源的外泌体YB-1/miR-223轴与神经细胞损伤之间的关联。免疫荧光,RT-PCR,ELISA和伤口愈合测定。
结果:这里,我们报道AD模型是导致小胶质细胞M1样(促炎)极化的原因,而后者又诱导神经细胞损伤.而M2样(抗炎)小胶质细胞可以释放富含miR-223的EXO,从而在体内和体外减轻AD模型中的神经炎症并改善神经损伤。此外,YB-1在细胞和EXO中直接与miR-223相互作用,并参与小胶质细胞外泌体miR-223的装载。
结论:这些结果表明,抗炎小胶质细胞介导的神经保护形成炎性损伤涉及通过YB-1分类的EXO输出miR-223。因此,YB-1介导的小胶质细胞外泌体分选miR-223促进神经细胞损伤修复,代表了AD的有希望的治疗目标。