IFN-γ刺激的MSCs重构Kupffer细胞表型并抑制肝细胞焦亡治疗急性肝衰竭的作用与机制研究

Previous studies have confirmed the effectiveness of MSCs transplantation for ALF, but the efficiency is limited and the mechanism is controversial. Recent studies indicate the dual-directional regulation effect between MSCs and immune microenvironment. This study aims to firstly improve the efficacy of MSCs for ALF through the appropriate pretreatment, then to explore the molecular mechanism of MSCs for ALF. Combined with our previous experiments and the latest research results, we hypothesize that IFN-γ pretreated MSCs attenuate acute liver failure via reprogramming of host kupffer cells to inhibit pyroptosis of hepatocytes may be the new mechanism of MSCs for ALF. To confirm our hypothesis, we intend to firstly compare the effect of different pretreatment MSCs for ALF, then to explore the key cytokines secreted by IFN-γ pretreated MSCs which reprogrammed of host kupffer cells via protein chip, next to evaluate whether IL-10 mediated the inhibiting effect on hepatocytes pyroptosis using knockout mouse technology, and at last to search for molecular mechanism of IL-10 inhibiting hepatocytes pyroptosis. The research will offer new theories and approach for clinical stem cell therapy of ALF.

既往研究证实了MSCs移植治疗ALF的有效性，但是其疗效有限，作用机制存在争议。目前发现MSCs和免疫微环境之间存在双向调节作用方式。本研究拟先通过合适的预处理，提高MSCs治疗ALF的疗效，随后寻找MSCs通过调节免疫微环境治疗ALF的分子机制。结合前期实验与本领域最新研究成果，我们提出假设：IFN-γ刺激的MSCs通过重构Kupffer细胞表型、强化IL-10的分泌继而抑制肝细胞焦亡或是其发挥肝保护作用的新机制。为证实以上猜想，我们拟：比较不同预处理方式MSCs治疗ALF的疗效；通过蛋白芯片寻找IFN-γ刺激的MSCs促使Kupffer细胞发生M2型极化的关键细胞因子；采用基因敲除小鼠技术探索IL-10是否介导了M2型Kupffer细胞抑制肝细胞的焦亡的过程；最后寻找IL-10抑制肝细胞的焦亡的分子机制。我们期望通过以上努力提高MSCs治疗ALF的疗效，丰富MSCs治疗ALF的理论。

MSCs移植已被证实在各种疾病中发挥积极作用。在本项目研究中证实：MSCs移植通过可溶性细胞因子及小分子生物活性物质的分泌参与到急性肝衰竭，部分肝切除及肝脏肿瘤模型等多种肝脏疾病当中。本课题通过文献调研，动物模型及体外实验的验证，证实MSCs移植可通过释放抗炎因子IL-10，降低NLRP3-caspase1炎症小体水平，减少急性肝衰竭小鼠模型中肝细胞焦亡；同时MSCs移植可通过PGE2的分泌，激活下游YAP的表达，进一步通过miR-29a-3p抑制PTEN激活mTOR通路，促进急性肝衰竭模型中肝细胞再生。进一步研究发现，MSCs移植对于大鼠部分肝切除模型也有治疗作用，MSCs移植通过mTOR信号改善肝脏脂质积累，MSCs的输注同时激活IL-6和YAP通路，表明MSCs有改善肝脏再生能力的潜力。在肿瘤模型中，我们也证实了低氧诱导的MSCs通过COX2/PGE2轴促进HCC的进展。本课题从多个模型中证实了MSCs强大的分泌能力及调节能力，证实了MSCs移植治疗临床应用的广泛前景。项目资助发表SCI论文4篇，培养博士研究生3人，硕士研究生4人。