基于p62-Keap1-Nrf2介导的自噬-氧化应激途径研究健脾清化颗粒改善NERD食管黏膜屏障损伤机制

食管黏膜屏障功能受损、通透性增加是NERD的重要病理基础，氧化应激反应是介导食管黏膜屏障损伤的主要因素。自噬是机体应对氧化应激损伤的重要途径之一，自噬流标志蛋白p62可激活Keap1-Nrf2通路，发挥抗氧化应激作用。前期研究结果表明，健脾清化颗粒可通过激活Keap1-Nrf2通路，减轻NERD食管黏膜屏障炎症性损伤，但其能否通过上调p62表达在上游发挥对该通路的激活作用有待进一步研究。本项目以NERD脾虚湿热证大鼠和体外食管上皮细胞损伤模型为研究对象，以p62调控Keap1-Nrf2信号通路为切入点，采用siRNA干扰、免疫荧光共聚焦、食管上皮跨膜电阻测定等检测手段，探讨健脾清化颗粒修复NERD食管黏膜屏障损伤的分子机制。本研究可进一步揭示健脾清化颗粒治疗NERD食管黏膜屏障损伤的重要靶标。

Esophageal mucosal barrier dysfunction and increased esophageal permeability are critical pathological basis of NERD. Oxidative stress is the main factor that mediates esophageal mucosal barrier damage. Autophagy is one of the important ways to cope with oxidative stress damage. Autophagic flux marker protein p62 can activate the Keap1-Nrf2 pathway and play an anti-oxidative stress effect. Previous studies have shown that Jianpi Qinghua granule can reduce the esophageal mucosal barrier inflammatory damage of NERD by activating the Keap1-Nrf2 pathway. However, further studies are still needed to clarify whether the granule can activate this pathway by up-regulating p62. In this study, the murine model of NERD with spleen deficiency and damp-heat syndrome as well as the model of esophageal epithelial injury in vitro will be studied to explore the molecular mechanism of Jianpi Qinghua granule in repairing the esophageal epithelial mechanical barrier dysfunction by regulating p62 to target Keap1-Nrf2 signaling pathway. SiRNA (small interfering RNA), immunofluorescence, mini-ussing chamber and other detection techniques will be used in this study. The study can further reveal the important target of Jianpi Qinghua granule in the treatment of NERD esophageal mucosal barrier dysfunction.