人经血干细胞来源外泌体包含的MCP-1改善肝纤维化的作用及机制研究

Liver fibrosis is a common pathway and common feature of various chronic liver diseases. Studies showed that menstrual blood-derived stem cell (MenSC) can improve liver fibrosis, but the role of exosomes in MenSC in liver fibrosis and its underlying molecular mechanisms are still unclear. Our pre-experiment found that exosomes in MenSC can improve liver fibrosis by inhibiting hepatic stellate cells (HSC), and high expression of MCP-1 was identified by the protein array of exosomes. Further in vitro cell experiments confirmed that MCP-1 also significantly inhibited the activation of the HSC. Therefore, we propose a hypothesis that "exosomes in MenSC may improve liver fibrosis by secreting MCP-1 targeting HSC". Based on this hypothesis, this project intends to use the cell model in vitro and mouse model of liver fibrosis to identify the therapeutic function and potential pathway of MCP-1 secreted by exosomes from MenSC-derived exosomes through biochemical and multi-group analysis. Through the research of this subject, we can further enrich the in-depth understanding exosomes in MenSC to improve liver fibrosis, and lay a solid theoretical foundation and realistic basis for the future application of cell-free treatment in liver fibrosis.

肝纤维化是各类慢性肝病的普遍途径和共同特征。已有研究证明人经血干细胞（MenSC）能够改善肝纤维化，但MenSC来源的外泌体（exosomes）对于肝纤维化的作用及其潜在分子机制尚不清楚。前期预实验发现MenSC中的exosomes能通过抑制肝星状细胞（HSC）改善肝纤维化，且exosomes蛋白芯片鉴定到高表达的MCP-1可显著地抑制HSC的激活和增殖。因此，本课题组提出“MenSC来源的exosomes可能通过包含的MCP-1抑制HSC改善肝纤维化”这一研究假说。基于该假说，本项目拟利用体外细胞模型、小鼠肝纤维化模型，通过分子生物学、细胞生物学和多组学分析，鉴定MenSC来源exosomes包含的MCP-1对肝纤维化的缓解功能以及潜在的作用机制。通过该研究，本课题组对MenSC中exosomes改善肝纤维化有更深入了解，为将来的无细胞治疗应用于肝纤维化提供理论支撑。

肝纤维化是各类慢性肝病的必经途径，如何有效地抑制肝纤维化是当前亟待解决的难题。已有研究证明人经血干细胞（又称宫血间充质干细胞，MenSC）能够改善肝纤维化，但MenSC来源的外泌体（exosomes）对于肝纤维化的作用及其潜在分子机制尚不清楚。基于此，本研究探讨了MenSC来源的exosomes（Exo）改善肝纤维化的可行性，更进一步阐述了MenSC-Exo缓解肝纤维化的内在分子机理和细胞机制。该项目主要研究了MenSC以及MenSC-Exo的特性鉴定，体内研究MenSC来源的exosomes中MCP-1在小鼠肝纤维化模型中的治疗效果及作用机制，体外研究MenSC来源的exosomes中抑制肝星状细胞（HSC）的作用和分子机制。通过该项目的开展，证实了OE-MCP-1 MenSC-Exo和MenSC-Exo移植到肝纤维化模型小鼠后的作用效果，揭示了MCP-1在缓解肝纤维化过程中的重要作用。同时体外通过MenSC-Exo和SiMCP-1-MenSC-Exo以及 transwell体外共培养表明了MenSC-Exo对LX-2细胞的增殖抑制作用，进一步阐明了MenSC-Exo中的MCP-1改善肝纤维化的重要作用。总之，该项目阐明了MenSC-Exo可能通过MCP-1抑制HSC改善肝纤维化的作用方式和分子机理，进一步丰富了MenSC来源的exosomes在肝病中的治疗作用，为肝纤维治疗提供一种潜在的治疗方式。