Bone marrow mesenchymal stem cell-derived exosomal microRNA-124-3p attenuates neurological damage in spinal cord ischemia-reperfusion injury by downregulating Ern1 and promoting M2 macrophage polarization
Bone marrow mesenchymal stem cell-derived exosomal microRNA-124-3p attenuates neurological damage in spinal cord ischemia-reperfusion injury by downregulating Ern1 and promoting M2 macrophage polarization
复制标题
骨髓间充质干细胞来源的外体microRNA-124-3p通过下调Ern1和促进M2巨噬细胞极化减轻脊髓缺血再灌注损伤的神经损伤
DOI：
10.1186/s13075-020-2146-x
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发表时间：
2020-04-09
影响因子：
4.9
通讯作者：
Yin, Fei
中科院分区：
文献类型：
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作者：
Li, Ran;Zhao, Kunchi;Yin, Fei
脊髓缺血再灌注损伤（SCIRI）常导致神经损伤和死亡。因此，了解SCIRI的病理，预防其发展具有重要的临床价值。方法分析骨髓间充质干细胞（BMMSC）来源的外泌体microRNA (miR)-124-3p在SCIRI中的作用。建立SCIRI大鼠模型，采用免疫组化、免疫荧光、RT-qPCR、western blot等方法检测巨噬细胞极化标志物Arg1、Ym1、Fizz中Ern1、M2的表达。通过生物信息学分析预测miR-124-3p与Ern1的靶向关系，并通过双荧光素酶报告基因试验验证。巨噬细胞与含有mir -124-3p的bmmsc来源的外泌体共培养。流式细胞术检测M2巨噬细胞，检测Arg1、Ym1、Fizz的表达。此外，给SCIRI大鼠注射含mir -124-3p的外泌体，TUNEL法观察脊髓细胞凋亡，H&E染色评价病理情况。结果SCIRI中，Ern1高表达，M2极化标记物低表达。沉默Ern1导致M2极化标记物的表达升高。MiR-124-3p靶向并负调控Ern1。外泌体miR-124-3p增强M2极化。高表达的外泌体miR-124-3p阻碍细胞凋亡，减轻scii诱导的组织损伤和神经损伤。来自bmmsc来源的外泌体的miR-124-3p通过抑制Ern1和促进M2极化改善SCIRI及其相关的神经损伤。综上所述，BMMSCs来源的外泌体miR-124-3p通过调节Ern1和M2巨噬细胞极化来减轻SCIRI诱导的神经损伤。
Background Spinal cord ischemia-reperfusion injury (SCIRI) often leads to neurological damage and mortality. In this regard, understanding the pathology of SCIRI and preventing its development are of great clinic value. Methods Herein, we analyzed the role of bone marrow mesenchymal stem cell (BMMSC)-derived exosomal microRNA (miR)-124-3p in SCIRI. A SCIRI rat model was established, and the expression of Ern1 and M2 macrophage polarization markers (Arg1, Ym1, and Fizz) was determined using immunohistochemistry, immunofluorescence assay, RT-qPCR, and western blot analysis. Targeting relationship between miR-124-3p and Ern1 was predicted using bioinformatic analysis and verified by dual-luciferase reporter assay. Macrophages were co-cultured with miR-124-3p-containing BMMSC-derived exosomes. M2 macrophages were identified using flow cytometry, and the expression of Arg1, Ym1, and Fizz was determined. In addition, SCIRI rats were injected with miR-124-3p-containing exosomes, spinal cord cell apoptosis was observed using TUNEL assay, and the pathological condition was evaluated with H&E staining. Results In SCIRI, Ern1 was highly expressed and M2 polarization markers were poorly expressed. Silencing Ern1 led to elevated expression of M2 polarization markers. MiR-124-3p targeted and negatively regulated Ern1. Exosomal miR-124-3p enhanced M2 polarization. Highly expressed exosomal miR-124-3p impeded cell apoptosis and attenuated SCIRI-induced tissue impairment and nerve injury. miR-124-3p from BMMSC-derived exosomes ameliorated SCIRI and its associated nerve injury through inhibiting Ern1 and promoting M2 polarization. Conclusion In summary, exosomal miR-124-3p derived from BMMSCs attenuated nerve injury induced by SCIRI by regulating Ern1 and M2 macrophage polarization.