CREG调控Kupffer细胞中NLRP3炎症小体活化在肝脏缺血再灌注损伤中的作用和机制研究

Hepatic ischemia-reperfusion injury (IRI) is one of the main constraints of clinical liver transplantation. Until now, there are no effective prevention measures. Kupffer cells are the key cells for amplifying liver damage during IRI. The NLRP3 inflammasome is the most important part of Kupffer cell activation. Our previous study found that the expression of E1A-activated gene repressor (CREG) was down-regulated in Kupffer cells during hypoxia-reoxygenation, and inhibition of CREG significantly induced the expression of NLRP3 inflammasome. This project is to investigate the biological function and mechanism of CREG in Kupffer cells during liver IRI: 1. Analyze the effect of CREG in Kupffer cells during the progression of liver IRI; 2. Determine the inhibitory effect of CREG on NLRP3 inflammasome in Kupffer cells; 3. In-depth study of the intrinsic mechanism of CREG in inhibiting the activation of NLRP3 inflammasome. This project is expected to elucidate the biological function of CREG in Kupffer cells during liver IRI and provide a new basis for clinical treatment of liver IRI.

肝脏缺血再灌注损伤（IRI）是临床上限制肝移植成功的主要制约因素之一，目前尚无确切有效的防治措施。Kupffer细胞是在IRI过程中放大肝脏损伤的关键细胞，其中NLRP3炎症小体是参与Kupffer细胞活化的重要环节。我们前期研究发现Kupffer细胞在缺氧复氧时E1A激活基因阻遏子（CREG）表达下调，且敲除巨噬细胞RAW264.7中的CREG后，NLRP3炎症小体的表达显著上调。本课题拟探讨Kupffer细胞中的CREG在肝脏IRI中的生物学功能及机制: ①利用小鼠IRI疾病模型分析Kupffer细胞中CREG在IRI中的作用；②明确CREG对Kupffer细胞中NLRP3炎症小体的抑制作用；③深入探讨CREG抑制NLRP3炎症小体的内在机制。本课题有望阐明Kupffer细胞中CREG在肝脏IRI的生物学功能，并为肝脏IRI的临床治疗提供新的依据。

肝脏缺血再灌注损伤（IRI）是临床上限制肝移植成功的主要制约因素之一，目前尚无确切有效的防治措施。Kupffer细胞（KC）是在IRI过程中放大肝脏损伤的关键细胞，其中NLRP3炎症小体是参与Kupffer细胞活化的重要环节。我们首先探讨了KC中CREG对肝脏IRI进展的影响，通过原代KC单独或与肝细胞共培养，观察敲除CREG前后对KC细胞以及肝细胞炎症因子的分泌，细胞增殖和细胞死亡；其次，研究了CREG对NLRP3炎症小体的调控作用，原代KC单独或与肝细胞共培养，观察敲除CREG前后对KC以及细胞炎症小体，下游相关分子和焦亡情况；再次，评估了NLRP3炎症小体对肝脏IRI进展的作用，利用原代CREG-的KC单独或与肝细胞共培养，观察CREG-的KC中NLRP3敲除或使用NLRP3抑制剂前后KC以及肝细胞炎症小体，下游相关分子和焦亡相关分子表达；最后探讨CREG是否通过TAK1/ASK1抑制KC中NLRP3炎症小体，验证CREG与TAK1/ASK1的互作，阻断KC中两者互作，检测阻断前后KC与肝细胞炎症小体，炎症因子，细胞焦亡的表达。本课题有望阐明Kupffer细胞中CREG在肝脏IRI的生物学功能，并为肝脏IRI的临床治疗提供新的依据。