BMSCs来源的外泌体miR-515-5p可下调TLR4/NLRP3通路抑制RA-FLS  

BMSCs-derived exosome miR-515-5p down-regulates the TLR4/NLRP3 pathway to inhibit synovioblast-like cells

作　　者：蔡东峰 钟超 杨子肖 李继模 张靖 洪嵩 Cai Dongfeng;Zhong Chao;Yang Zixiao;Li Jimo;Zhang Jing;Hong Song(Department of Orthopedics,Affiliated Hospital of Zunyi Medical University,Zunyi Guizhou 563000,China;Department of Orthopedics,Bijie Hospital,Zhejiang Provincial People’s Hospital,Bijie Guizhou 551700,China)

机构地区：[1]遵义医科大学附属医院骨科,贵州遵义563000 [2]浙江省人民医院毕节医院骨科,贵州毕节551700

出　　处：《遵义医科大学学报》2025年第3期252-262,269,共12页Journal of Zunyi Medical University

基　　金：贵州省科技厅基础研究计划项目[NO:黔科合基础-ZK(2023)一般542];贵州省卫健委科技基金资助项目(NO:gzwjk2020-1-127)。

摘　　要：目的 探讨骨髓间充质干细胞(BMSCs)外泌体miR-515-5p介导TLR4/NLRP3通路对类风湿性关节炎滑膜成纤维样细胞(RA-FLS)增殖、凋亡、迁移和侵袭的影响。方法 体外培养BMSCs、RA-FLS细胞,制备miR-515-5p低表达的BMSCs及si-TLR4转染的RA-FLS细胞。分为4组:RA-FLS细胞组、RA-FLS细胞+外泌体组,RA-FLS细胞+miR-515-5p inhibitor外泌体组、si-TLR4转染RA-FLS细胞+miR-515-5p inhibitor外泌体组。依次对各组RA-FLS行EDU、CCK-8、流式凋亡检测、划痕实验、Transwell、ELISA及Western blot检测。结果 BMSCs外泌体组中,miR-515-5p可明显抑制RA-FLS细胞增殖并促进其凋亡,抑制其侵袭、迁移及释放IL-6、TNF-α、IL-1β炎性因子,抑制TLR4、NLRP3蛋白表达。抑制miR-515-5p后,RA-FLS细胞的TLR4、NLRP3蛋白表达上调,但对si-TLR4转染的RA-FLS细胞,其TLR4、NLRP3蛋白表达无上调。结论 BMSCs细胞来源的外泌体miR-515-5p通过靶向下调TLR4/NLRP3通路而抑制RA-FLS细胞的增殖、迁移、侵袭和炎症因子释放并促进其凋亡。Objective To explore the effect of the TLR4/NLRP3 pathway mediated by exosome miR-515-5p of bone marrow mesenchymal stem cells(BMSCs)on the proliferation,apoptosis,migration,and invasion of synovioblast-like cells(RA-FLS)in rheumatoid arthritis.Methods BMSCs and RA-FLS cells were cultured in vitro to prepare BMSCs with low expression of miR-515-5p and RA-FLS cells transfected with si-TLR4.RA-FLS cells and transfected cells were divided into 4 groups:RA-FLS control,RA-FLS+exosome treatment group,RA-FLS+miR-515-5p inhibitor exosome treatment group,si-TLR4 transfected RA-FLS cells+miR-515-5p inhibitor exosome treatment group.EDU,CCK-8,flow apoptosis,scratch test,Transwell,ELISA and Western blot were performed to detect biological changes.Results In the BMSCs exosome group,miR-515-5p significantly inhibited the proliferation and apoptosis of RA-FLS cells,suppressed their invasion,migration,and the release of inflammatory cytokines such as IL-6,TNF-α,and IL-1β,and downregulated the expression of TLR4 and NLRP3 proteins.Upon inhibition of miR-515-5p,the expression levels of TLR4 and NLRP3 proteins in RA-FLS cells were upregulated;however,this upregulation is not observed in RA-FLS cells transfected with Si-TLR4.Conclusion BMSCs cell-derived exosome miR-515-5p inhibits the proliferation,migration,invasion and release of inflammatory factors of RA-FLS cells,and promotes their apoptosis by targeting down-regulation of the TLR4/NLRP3 pathway.

关 键 词：骨髓间充质干细胞 外泌体 miR-515-5p TLR4 NLRP3 RA-FLS 

分 类 号：R684.3[医药卫生—骨科学]