Li Feng, Yan Tengfei, Wang Shunlan, Wen Xiaohong
Department of Clinical Laboratory, Hainan General Hospital (Hainan Affiliated Hospital of Hainan Medical University), Haikou, 570311, Hainan, China.
Baoding First Central Hospital, Baoding, 071000, Heibei, China.
Clin Exp Med. 2023 Nov;23(7):4021-4031. doi: 10.1007/s10238-023-01122-0. Epub 2023 Jun 24.
Liver fibrosis is a serious stage of chronic liver injury. Inhibition of hepatic stellate cells activation and hepatocytes apoptosis is important measures in the treatment of liver fibrosis. Studies have shown that exosomes are involved in regulating the information transmission between cells, but there are few studies on the interaction between exosomes from HSC and hepatocytes. This study screened miRNAs with significant differences related to liver fibrosis in the database. Then, we activated HSC applying transforming growth factor β1 (TGF-β1) and collected exosomes. The expression of miRNA in HSC-derived exosomes was verified by quantitative real-time PCR (qRT-PCR). The results of cell function test showed that HSC-derived exocrine miRNA-99a-5p could inhibit hepatocytes proliferation and promote hepatocytes apoptosis. Conversely, inhibition of miRNA-99a-5p can promote hepatocytes proliferation and inhibit apoptosis. Target gene prediction and luciferase assay show that miRNA can specifically bind to BMPR2 site sequence. In addition, we also detected the expression of BMPR2 and apoptosis-related protein by qRT-PCR and Western blot (WB). In conclusion, this study demonstrates that HSC-derived exocrine miRNA-99a-5p can promote hepatocytes apoptosis and participate in the process of liver fibrosis by targeting BMPR2. Our findings highlight the therapeutic potential of HSC-derived exocrine miRNA-99a-5p in hepatic fibrosis.
肝纤维化是慢性肝损伤的严重阶段。抑制肝星状细胞活化和肝细胞凋亡是治疗肝纤维化的重要措施。研究表明，外泌体参与调节细胞间的信息传递，但关于肝星状细胞来源的外泌体与肝细胞之间相互作用的研究较少。本研究在数据库中筛选出与肝纤维化相关的具有显著差异的微小RNA（miRNA）。然后，我们应用转化生长因子β1（TGF-β1）激活肝星状细胞并收集外泌体。通过定量实时聚合酶链反应（qRT-PCR）验证肝星状细胞来源的外泌体中miRNA的表达。细胞功能测试结果表明，肝星状细胞来源的外分泌miRNA-99a-5p可抑制肝细胞增殖并促进肝细胞凋亡。相反，抑制miRNA-99a-5p可促进肝细胞增殖并抑制凋亡。靶基因预测和荧光素酶测定表明，miRNA可特异性结合骨形态发生蛋白受体2（BMPR2）位点序列。此外，我们还通过qRT-PCR和蛋白质免疫印迹法（WB）检测了BMPR2及凋亡相关蛋白的表达。总之，本研究表明，肝星状细胞来源的外分泌miRNA-99a-5p可通过靶向BMPR2促进肝细胞凋亡并参与肝纤维化进程。我们的研究结果突出了肝星状细胞来源的外分泌miRNA-99a-5p在肝纤维化治疗中的潜力。
