Wang Xiujuan, Yang Ying, Li Wei, Hao MingYuan, Xu YongSheng
Technology Department, Everunion Biotechnology Co. LTD, Tianjin, China.
Biochem Biophys Rep. 2023 Jul 20;35:101497. doi: 10.1016/j.bbrep.2023.101497. eCollection 2023 Sep.
Spinal cord injury (SCI) is an incurable central nervous system impairment that lack of efficient treatment. Exosomes derived from mesenchymal stem cells (MSCs) are widely applied in disease treatment. This work aimed to determine the promising therapeutic effects of MSC-derived exosomal miRNA146b on SCI. A rat spinal cord injury (SCI) model and lipopolysaccharide (LPS)-induced PC12 cell model were established. Exosomes were extracted from human umbilical cord mesenchymal stem cells (hUCMSCs). The identification of exosomes was performed by using transmission electronic microscope (TEM) and nanoparticle tracking analysis (NTA). Hematoxylin and eosin (HE) staining and TUNEL assay were performed to assess tissue damage and apoptosis, respectively. ELISA was performed to detect levels of inflammatory cytokines. Cell viability was checked by cell counting kit 8 (CCK-8). Gene expression and protein levels were detected by qPCR and western blotting assay. The interaction between miR-146 b and Toll-like receptor 4 (TLR4) was assessed by luciferase reporter gene assay. The hUCMSC-derived exosomes could notably alleviate the spinal cord injury and cell apoptosis. The exosomal miR-146 b treatment suppressed the release of IL-1 β, IL-6, and TNFα. The miR-146 b suppressed the expression of TLR4, directly interact with the 3'-untranslated region (3'UTR) of TLR4, and inactivated the nuclear factor κB (NF-κB) signaling. The hUCMSCs-derived exosomal miR-146 b protects neurons from spinal cord injury through targeting the TLR4 and inactivating the NF-κB signaling. Our findings supported the application of hUCMSCs-derived exosomal miR-146 b for the protection of SCI.
脊髓损伤（SCI）是一种无法治愈的中枢神经系统损伤，缺乏有效的治疗方法。间充质干细胞（MSCs）来源的外泌体被广泛应用于疾病治疗。本研究旨在确定MSCs来源的外泌体miRNA146b对SCI的潜在治疗作用。建立了大鼠脊髓损伤（SCI）模型和脂多糖（LPS）诱导的PC12细胞模型。从人脐带间充质干细胞（hUCMSCs）中提取外泌体。使用透射电子显微镜（TEM）和纳米颗粒跟踪分析（NTA）对外泌体进行鉴定。分别进行苏木精-伊红（HE）染色和TUNEL检测以评估组织损伤和细胞凋亡。通过酶联免疫吸附测定（ELISA）检测炎性细胞因子水平。使用细胞计数试剂盒8（CCK-8）检测细胞活力。通过qPCR和蛋白质印迹法检测基因表达和蛋白质水平。通过荧光素酶报告基因检测评估miR-146b与Toll样受体4（TLR4）之间的相互作用。hUCMSC来源的外泌体可显著减轻脊髓损伤和细胞凋亡。外泌体miR-146b治疗抑制了IL-1β、IL-6和TNFα的释放。miR-146b抑制TLR4的表达，直接与TLR4的3'-非翻译区（3'UTR）相互作用，并使核因子κB（NF-κB）信号失活。hUCMSCs来源的外泌体miR-146b通过靶向TLR4并使NF-κB信号失活来保护神经元免受脊髓损伤。我们的研究结果支持hUCMSCs来源的外泌体miR-146b在保护SCI方面的应用。
