Department of Gastroenterology, Wuhan Third Hospital, Tongren Hospital of Wuhan University, Wuhan, China.
J Histotechnol. 2023 Dec;46(4):158-169. doi: 10.1080/01478885.2023.2215151. Epub 2023 May 25.
Bone marrow mesenchymal stem cell (BSMC)-derived extracellular vehicles (EVs) have a pivotal therapeutic potential in hepatic fibrosis (HF). Activation of hepatic stellate cells (HSCs) is the key mechanism in HF progression. Downregulation of miR-192-5p was previously observed in activated HSCs. Nonetheless, the functions of BSMC-derived exosomal miR-192-5p in activated HSCs remain unclear. In this study, transforming growth factor (TGF)-β1 was used to activate HSC-T6 cells to mimic HF in vitro. Characterization of BMSCs and BMSC-derived EVs was performed. Cell-counting kit-8 assay, flow cytometry, and western blotting revealed that TGF-β1 increased cell viability, promoted cell cycle progression, and induced upregulation of fibrosis markers in HSC-T6 cells. Overexpression of miR-192-5p or BMSC-derived exosomal miR-192-5p suppressed TGF-β1-triggered HSC-T6 cell activation. RT-qPCR revealed that protein phosphatase 2 regulatory subunit B'' alpha (PPP2R3A) was downregulated in miR-192-5p-overexpressed HSC-T6 cells. Luciferase reporter assay was used for verifying the relation between miR-192-5p and PPP2R3A, which showed that miR-192-5p targeted PPP2R3A in activated HSC-T6 cells. Collectively, BMSC-derived exosomal miR-192-5p targets PPP2R3A and inhibits activation of HSC-T6 cells.
骨髓间充质干细胞(BSMC)衍生的细胞外囊泡(EVs)在肝纤维化(HF)中具有重要的治疗潜力。肝星状细胞(HSCs)的激活是 HF 进展的关键机制。先前在激活的 HSCs 中观察到 miR-192-5p 的下调。然而,BSMC 衍生的外泌体 miR-192-5p 在激活的 HSCs 中的功能仍不清楚。在本研究中,使用转化生长因子(TGF)-β1 激活 HSC-T6 细胞以模拟体外 HF。对 BMSC 和 BMSC 衍生的 EV 进行了表征。细胞计数试剂盒-8 测定、流式细胞术和 Western blot 揭示 TGF-β1 增加了细胞活力,促进了细胞周期进程,并诱导了 HSC-T6 细胞中纤维化标志物的上调。miR-192-5p 的过表达或 BMSC 衍生的外泌体 miR-192-5p 抑制了 TGF-β1 触发的 HSC-T6 细胞激活。RT-qPCR 显示 miR-192-5p 过表达的 HSC-T6 细胞中蛋白磷酸酶 2 调节亚基 B''alpha(PPP2R3A)下调。荧光素酶报告基因测定用于验证 miR-192-5p 和 PPP2R3A 之间的关系,结果表明 miR-192-5p 在激活的 HSC-T6 细胞中靶向 PPP2R3A。综上所述,BSMC 衍生的外泌体 miR-192-5p 靶向 PPP2R3A 并抑制 HSC-T6 细胞的激活。