靶向阻断EPAS1基因调控TGF-β/Smad4信号通路在妊娠期SAP肠屏障损伤中的作用机制研究

随着我国计划生育政策的调整及高龄孕妇的增加，妊娠期重症急性胰腺炎（SAP）的发病率也明显上升。EPAS1是新近发现的一个含有ODD结构域的细胞因子，是机体炎症/缺氧微环境中最重要的一员，我们的前期研究也发现：EPAS1在SAP肠粘膜屏障损伤中起重要作用，但其作用机制还有待进一步研究。基于此，本项目利用生物纳米技术和基因靶向技术相融合，构建靶向EPAS1基因的白蛋白纳米微球实现高效、缓释，采用qRT-PCR、ELISA、FCM等经典方法，从分子、细胞、动物模型多水平探讨EPAS1基因对妊娠期SAP肠粘膜上皮细胞增殖、凋亡的调控，揭示EPAS1通过抑制TGF-β表达，从而下调TGF-β/Smad4信号通路下游靶基因miR-130a、Occludin等表达的分子机制，明确靶向EPAS1基因的纳米微球对肠屏障损伤的治疗效果，本研究将为妊娠期SAP肠屏障损伤中靶基因选择和基因载体的优化提供新的策略。

Severe Acute Pancreatitis (SAP) in pregnancy is a disease of rapid progression, high mortality rate, and complicated pathogenesis. The incidence of SAP in China were increasing significantly with the adjustment of China's family planning policy as well as the amount of women of advanced maternal age. EPAS1 is a newly found cytokin with ODD domain as well as a most important component in the inflammatory/hypoxic microenvironment of the human body. Our previous study indicated that EPAS1 plays a pivotal role in the SAP-induced intestinal barrier dysfunction, while the mechanism remains to be illuminated. On this basis, the gene-nano composites was prepared by the combination of bio-nanotechnology and gene targeting technology. qRT-PCR, WB, FCM and other classical methods were applied to explore the role of EPAS1 in regulating Crosstalk between inflammatory/hypoxic microenvironment and intestinal mucosal barrier via TGF -β/Smad4 signal pathway. The results revealed that EPAS1 regulate proliferation and apoptosis of intestinal mucosa epithelial cells through down regulating the expression of Occludin, miR-130a and other downstream target genes of TGF-β/Smads pathway, so as to provide a new potential strategy for the treatment of SAP intestinal mucosal barrier dysfunction during pregnancy.