Umbilical mesenchymal stem cell-derived exosomes facilitate spinal cord functional recovery through the miR-199a-3p/145-5p-mediated NGF/TrkA signaling pathway in rats.
Umbilical mesenchymal stem cell-derived exosomes facilitate spinal cord functional recovery through the miR-199a-3p/145-5p-mediated NGF/TrkA signaling pathway in rats.
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脐带间充质干细胞来源的外泌体通过 miR-199a-3p/145-5p 介导的 NGF/TrkA 信号通路促进大鼠脊髓功能恢复
DOI：
10.1186/s13287-021-02148-5
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发表时间：
2021-02-12
影响因子：
7.5
通讯作者：
Rong L
中科院分区：
文献类型：
--
作者：
Wang Y;Lai X;Wu D;Liu B;Wang N;Rong L
尽管作为人脐带间充质干细胞(Huc-MSCs)的副产物，Exosome已被证明是治疗创伤性脊髓损伤(SCI)的有效药物，但其作用机制尚不清楚。方法我们设计并进行了这项研究，以确定Exosome是否通过减轻继发性炎症风暴引起的神经元损伤和促进轴突生长来减轻SCI的损伤。结果miR-199a-3p/145-5p是外切体中表达相对较高的miRNAs，可通过调节NGF/TrkA途径促进脂多糖抑制PC12细胞的体外分化。我们还证明了Cblb是miR-199a-3p的直接靶标，Cb1是miR-145-5p的直接靶标。Cblb和Cbl基因敲除导致TrkA泛素化水平显著降低，随后激活了NGF/TrkA下游通路Akt和Erk。相反，Cblb和Cbl的过表达与TrkA泛素化水平显著增加相关，随后使NGF/TrkA下游通路Akt和Erk失活。Western印迹和免疫共沉淀实验证实TrkA与Cblb、TrkA与Cb1之间存在直接相互作用。体内实验发现，外切体miR-199a-3p/145-5p可上调损伤部位TrkA的表达，并促进脊髓损伤大鼠的运动功能。结论综上所述，将miR-199a-3p/145-5p导入脊髓损伤大鼠神经元的外切体能影响TrkA泛素化，促进NGF/TrkA信号通路，提示Huc-MSC来源的外切体可能是一种很有前途的脊髓损伤治疗策略。
BackgroundAlthough exosomes, as byproducts of human umbilical cord mesenchymal stem cells (hUC-MSCs), have been demonstrated to be an effective therapy for traumatic spinal cord injury (SCI), their mechanism of action remains unclear.MethodsWe designed and performed this study to determine whether exosomes attenuate the lesion size of SCI by ameliorating neuronal injury induced by a secondary inflammatory storm and promoting neurite outgrowth. We determined the absolute levels of all exosomal miRNAs and investigated the potential mechanisms of action of miR-199a-3p/145-5p in inducing neurite outgrowth in vivo and in vitro.ResultsmiR-199a-3p/145-5p, which are relatively highly expressed miRNAs in exosomes, promoted PC12 cell differentiation suppressed by lipopolysaccharide (LPS) in vitro through modulation of the NGF/TrkA pathway. We also demonstrated that Cblb was a direct target of miR-199a-3p and that Cbl was a direct target of miR-145-5p. Cblb and Cbl gene knockdown resulted in significantly decreased TrkA ubiquitination levels, subsequently activating the NGF/TrkA downstream pathways Akt and Erk. Conversely, overexpression of Cblb and Cbl was associated with significantly increased TrkA ubiquitination level, subsequently inactivating the NGF/TrkA downstream pathways Akt and Erk. Western blot and coimmunoprecipitation assays confirmed the direct interaction between TrkA and Cblb and TrkA and Cbl. In an in vivo experiment, exosomal miR-199a-3p/145-5p was found to upregulate TrkA expression at the lesion site and also promote locomotor function in SCI rats.ConclusionsIn summary, our study showed that exosomes transferring miR-199a-3p/145-5p into neurons in SCI rats affected TrkA ubiquitination and promoted the NGF/TrkA signaling pathway, indicating that hUC-MSC-derived exosomes may be a promising treatment strategy for SCI.
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