Xu Huikang, Fu Jiamin, Chen Lijun, Zhou Sining, Fang Yangxin, Zhang Qi, Chen Xin, Yuan Li, Li Yifei, Xu Zhenyu, Xiang Charlie
State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310003, China.
Research Units of Infectious Disease and Microecology, Chinese Academy of Medical Sciences, Hangzhou, Zhejiang 310003, China.
Stem Cells Int. 2023 Feb 28;2023:2988907. doi: 10.1155/2023/2988907. eCollection 2023.
Human menstrual blood-derived mesenchymal stem cells (MenSCs) and their secreted small extracellular vesicles (EVs) had been proven to relieve inflammation, tissue damage, and fibrosis in various organs. The microenvironment induced by inflammatory cytokines can promote mesenchymal stem cells (MSCs) to secrete more substances (including EVs) that could regulate inflammation. Inflammatory bowel disease (IBD) is a chronic idiopathic intestinal inflammation, the etiology and mechanism of which are unclear. At present, the existing therapeutic methods are ineffective for many patients and have obvious side effects. Hence, we explored the role of tumor necrosis factor - (TNF--) pretreated MenSC-derived small EV (MenSCs-sEV ) in a mouse model of dextran sulfate sodium- (DSS-) induced colitis, expecting to find better therapeutic alterations. In this research, the small EVs of MenSCs were obtained by ultracentrifugation. MicroRNAs of small EVs derived from MenSCs before and after TNF- treatment were sequenced, and the differential microRNAs were analyzed by bioinformatics. The small EVs secreted by TNF--stimulating MenSCs were more effective in colonic mice than those secreted directly by MenSCs, as evidenced by the results of histopathology analysis of colonic tissue, immunohistochemistry for tight junction proteins, and enzyme-linked immunosorbent assay (ELISA) for cytokine expression profiles in vivo. The process of MenSCs-sEV relieving colonic inflammation was accompanied by the polarization of M2 macrophages in the colon and miR-24-3p upregulation in small EVs. In vitro, both MenSC-derived sEV (MenSCs-sEV) and MenSCs-sEV reduced the expression of proinflammatory cytokines, and MenSCs-sEV can increase the portion of M2 macrophages. In conclusion, after TNF- stimulation, the expression of miR-24-3p in small EVs derived from MenSCs was upregulated. MiR-24-3p was proved to target and downregulate interferon regulatory factor 1 (IRF1) expression in the murine colon and then promoted the polarization of M2 macrophages. The polarization of M2 macrophages in colonic tissues then reduced the damage caused by hyperinflammation.
人月经血来源的间充质干细胞（MenSCs）及其分泌的小细胞外囊泡（EVs）已被证明可缓解各种器官的炎症、组织损伤和纤维化。炎性细胞因子诱导的微环境可促进间充质干细胞（MSCs）分泌更多可调节炎症的物质（包括EVs）。炎症性肠病（IBD）是一种慢性特发性肠道炎症，其病因和机制尚不清楚。目前，现有的治疗方法对许多患者无效且有明显副作用。因此，我们在葡聚糖硫酸钠（DSS）诱导的结肠炎小鼠模型中探索了肿瘤坏死因子-（TNF-）预处理的MenSC来源的小EV（MenSCs-sEV）的作用，期望找到更好的治疗方法。在本研究中，通过超速离心获得MenSCs的小EVs。对TNF-处理前后MenSCs来源的小EVs的微小RNA进行测序，并通过生物信息学分析差异微小RNA。TNF-刺激的MenSCs分泌的小EVs在结肠小鼠中比MenSCs直接分泌的小EVs更有效，这在结肠组织的组织病理学分析、紧密连接蛋白的免疫组织化学以及体内细胞因子表达谱的酶联免疫吸附测定（ELISA）结果中得到了证实。MenSCs-sEV缓解结肠炎症的过程伴随着结肠中M2巨噬细胞的极化和小EVs中miR-24-3p的上调。在体外，MenSC来源的sEV（MenSCs-sEV）和MenSCs-sEV均降低了促炎细胞因子的表达，并且MenSCs-sEV可增加M2巨噬细胞的比例。总之，TNF-刺激后，MenSCs来源的小EVs中miR-24-3p的表达上调。已证明miR-24-3p靶向并下调小鼠结肠中干扰素调节因子1（IRF1）的表达，进而促进M2巨噬细胞的极化。结肠组织中M2巨噬细胞的极化随后减少了过度炎症引起的损伤。
