沉默lncRNA-HOTAIR 对重症急性胰腺炎小鼠肠黏膜屏障保护作用的实验研究

胰腺继发感染是重症急性胰腺炎（SAP）最严重的并发症,肠道是引发胰腺继发感染的主要来源。本课题组前期研究表明,体内炎症因子失控性释放是SAP肠黏膜屏障损伤的主要原因，最新研究发现lncRNAs与单核巨噬细胞炎症反应密切相关。我们的前期工作已证实lncRNAs在小鼠SAP模型中差异表达，且lncRNA-HOTAIR在SAP小鼠肠上皮中的表达明显升高。本项目拟体外转染lncRNA-HOTAIR siRNA至肠上皮细胞，并通过RNA-pulldown实验联合质谱分析，鉴定与lncRNA-HOTAIR相互作用的蛋白，探讨其对SAP炎症反应及肠屏障作用及机制，并在此基础上，将in vivo-jetPEITM介导的lncRNA-HOTAIR SiRNA静脉注入SAP小鼠模型，探讨其对SAP小鼠炎症反应的影响及肠黏膜屏障保护作用及其机制，为临床上基因干预SAP肠黏膜屏障损伤的可行性提供依据。

Pancreatic secondary infection is the most serious complication of severe acute pancreatitis (SAP), intestinal tract is the main origin to cause secondary infection. Our previous study showed that the uncontrolled release of inflammatory cytokines is the main reason to induce SAP-associated intestinal mucosal barrier damage in vivo, recently study finds lncRNAs take part in inflammation response led by monocytes and macrophage. Our previous work has demonstrated the expression of lncRNAs is different markedly in SAP mouse model and the expression of lncRNA-HOTAIR is significantly increased in the intestinal mucosal of SAP mouse model .the present project intends to transfect lncRNA- HOTAIR siRNA to intestinal epithelial cells in vitro, identify the protein which can interact with lncRNA-HOTAIR by RNA-pulldown experiments and mass spectrometry , explore the function and mechanism how lncRNA-HOTAIR influences SAP-associated inflammation and intestinal barrier damage. On this basis, we inject lncRNA-HOTAIR SiRNA mediated by in vivo-jetPEITM to vein of SAP mouse model, investigate the mechanism lncRNA-HOTAIR affects inflammation and protects intestinal mucosal barrier from damage in SAP mouse model , thus provide the evidence for the feasibility that clinical gene intervenes the SAP-associated intestinal mucosal barrier damage .