白藜芦醇诱导线粒体自噬对腹膜间皮细胞炎症的保护机制

Peritoneal dialysis is an important mode of renal replacement therapy for patients with end-stage renal disease due to it's preservation of residual renal function and initial survival advantage. Long-term application of traditional glucose-base peritoneal dialysis fluids can trigger chronic inflammation in peritoneal mesothelial cells, and launch angiogenesis and fibrosis. At present, the role of mitochondria in cells has aroused increased attention, and recent studies have demonstrated that timely launched mitophagy can remove damaged mitochondria, suppress the reactive oxygen species (ROS) generation and NLRP3 inflammasome activation and downregulate pro-inflammatory cytokines such as interleukin (IL-1β、IL-18), which can avoid the tissue lesion caused by excessively activated inflammatory response. This project is to investigate the mechanism of resveratrol-induced mitophagy through AMPK activation and the protective role of the inflammation caused by glucose-base peritoneal dialysis fluids in peritoneal mesothelial cells by the application of confocal fluorescence microscope, electron microscope, small interfering RNA（siRNA）and immunoblot. Our goal is to explore the new therapeutical target of inflammation in peritoneal cavity and fibrosis associated with peritoneal dialysis, which could have better clinical perspectives.

腹膜透析因其具有良好的残肾保护及早期生存率优势而成为终末期肾病患者的重要替代治疗方法，长期应用传统葡萄糖腹膜透析液可导致腹膜间皮细胞慢性炎症，启动新生血管形成及腹膜组织纤维化。当前，线粒体在细胞炎症反应中的作用备受关注，最近研究证实：适时启动线粒体自噬可清除细胞内受损线粒体，减少活性氧（ROS）的产生，降低NLRP3炎症体激活，下调处于炎症、免疫反应上游的主要效应因子IL-1β、IL-18的表达，避免过度炎症反应对机体组织的损伤。本项目以线粒体自噬为切入点，根据国际上最新研究进展，应用激光共聚焦显微镜、扫描电镜、siRNA、免疫印迹等方法，探讨白藜芦醇激活AMPK信号通路诱导线粒体自噬的机制以及对含糖腹透液所致腹膜间皮细胞炎症反应的保护机制，为腹膜透析腹腔局部炎症及腹膜组织纤维化寻找新的干预靶点提供可能重要的实验依据，具有较好的临床应用前景。

研究背景：腹膜透析因其具有良好的残肾保护及早期生存率优势而成为终末期肾病患者的重要替代治疗方法，长期应用传统葡萄糖腹膜透析液可导致腹膜间皮细胞慢性炎症。新近研究证实：适时启动线粒体自噬可清除细胞内受损线粒体，减少活性氧（ROS）的产生，降低NLRP3炎症体激活，下调处于炎症、免疫反应上游的主要效应因子IL-1β的表达，避免过度炎症反应对机体组织的损伤。.研究内容：人腹膜间皮细胞株（HMrSV5）作为研究对象。第一，探讨葡萄糖腹膜透析液诱导激活线粒体活性氧（ROS）-NLRP3炎症小体机制。第二，研究白藜芦醇诱导细胞自噬及机制。第三，探讨调节自噬对线粒体ROS-NLRP3-IL-1β的影响。.研究结果及意义：葡萄糖腹透液尤其是高糖腹透液作用人腹膜间皮细胞可激活线粒体ROS-NLRP3-IL-1β；另一方面，白藜芦醇可通过激活AMPK信号通过诱导线粒体自噬，负调节自噬可增加线粒体ROS，增强NLRP3激活；正调节自噬对线粒体ROS无影响，减弱NLRP3激活，提示白藜芦醇诱导人腹膜间皮细胞线粒体自噬减轻ROS-NLRP3介导的腹膜炎症损伤，有可能成为临床干预长期应用葡萄糖腹膜透析液所致腹膜炎症损伤的靶点。