目的：观察高糖腹膜透析液对人腹膜间皮细胞NLRP3-IL-1β的影响。方法：体外培养人腹膜间皮细胞株第5～10代［HMrSV5，DMEM/F12培养基含10%（体积分数）胎牛血清］，观察1.5%、2.5%、4.25%（质量分数）葡萄糖腹膜透析液（dextrose）及线粒体呼吸链复合酶Ⅰ抑制剂鱼藤酮（rotenone），线粒体呼吸链复合酶Ⅱ抑制剂噻吩甲酰三氟丙酮（thenoyltrifluoroacetone，TTFA），线粒体呼吸链复合酶Ⅲ抑制剂抗霉素A（antimycin A）对细胞IL-1β表达的影响（免疫印迹）；通过小RNA干扰技术下调NLRP3的表达，免疫印迹分析4.25%高糖腹膜透析液作用下细胞IL-1β的表达；通过白藜芦醇（resveratrol，RSV）诱导自噬，3-甲基腺嘌呤（3-methyladenine，3-MA）、自噬相关蛋白5（autophagy related gene 5，ATG5）siRNA、自噬相关蛋白（Beclin1）siRNA抑制自噬，流式细胞仪分析细胞活性氧（reactive oxygen species，ROS），免疫印迹分析IL1β的表达。结果：对照组、1.5%高糖腹膜透析液、2.5%高糖腹膜透析液、4.25%高糖腹膜透析液、鱼藤酮（10 μmol/L）、抗霉素A（50 mg/L）、噻吩甲酰三氟丙酮（10 μmol/L）各组细胞上清IL-1β的相对表达依次为0、0.175±0.082、0.418±0.163、2.357±0.288、2.642±0.358、3.271±0.462、0.123±0.091，提示 2.5%高糖腹膜透析液、4.25%高糖腹膜透析液、鱼藤酮、抗霉素A作用细胞IL-1β表达明显升高，应用NLRP3 siRNA可阻断上述效应；此外，对照组、阴性对照siRNA（negative control，NC siRNA）、RSV（50 μmol/L，诱导自噬）、3MA（10 mmol/L，抑制自噬）、ATG5 siRNA（抑制自噬）、Beclin1 siRNA（抑制自噬）各组总线粒体荧光强度分别为1.76±0.42、1.83±0.55、1.85±0.62、7.36±0.92、5.35±0.77、5.06±0.62，超氧化线粒体荧光强度分别为821.68±95.12、868.15±102.82、723.39±92.56、1 660.08±113.65、1 433.01±107.24、1 562.36±112.88，结合免疫印迹结果，提示抑制自噬可增强线粒体ROS产生和提高IL-1β表达，诱导自噬对ROS、IL-1β无明显影响。结论：长期应用高糖腹膜透析液，腹膜间皮细胞NLRP3-IL-1β持续激活，实时启动自噬可能成为干预NLRP3-IL-1β持续激活、延缓腹膜透析腹腔慢性炎症及腹膜纤维化的治疗靶点。

Objective： To explore the effect of high glucose-based peritoneal dialysis fluids on NLRP3-IL-1β in human peritoneal mesothelial cells. Methods: HMrSV5 cells (SV40 immortalized human peritoneal mesothelial cell line) were grown in type Ⅰ collagen-coated dishes in DMEM/F12 containing 10% fetal calf serum (FCS). All experiments on HMrSV5 cells were performed between passages 5 and 10-The cells were divided into 7 groups: control, 1-5% dextrose, 2-5% dextrose, 425% dextrose, rotenone, thenoyltrifluoroacetone (TTFA), and antimycin A-Immunoblotting was used to evaluate the expression of IL-1β-Small interfering RNA (siRNA) targeting NLRP3 was used to downregulate the expression of NLRP3 and Western blot was used to evaluate the expression of IL-1β in human peritoneal mesothelial cells exposed to 4-25% dextrose- In the meanwhile, resveratrol （RSV） was used to induce autophagy, 3-methyladenine (3-MA) and siRNA against Beclin 1 or ATG5 were used to block auto-phagy, flow cytometric was used to analyze the respiring (mitotracker deep red), total (mitotracker green) and reactive oxygen species (ROS)generating mitochondria (mitoSOX); Western blot was used to evaluate the expression of IL-1β- Results: The IL-1β relative expressions were 0, 0-175±0-082, 0-418±0-163, 2-357±0-288, 2-642±0-358, 3-271±0-462, and 0-123±0-091, indicating that the cells exposed to high glucose-based peritoneal dialysis fluids and cells treated with mitochondria respiratory chain key enzyme complex Ⅰ, and complex Ⅲ inhibitors increased the IL-1β expression- And we found that NLRP3 knock-down significantly blocked the upregulation of IL-1β- In addition, the fluorescence intensity of total mitochondria and ROS-generating mitochondria in the following groups: control, negative control, RSV, 3-MA, ATG5 siRNA, Beclin1 siRNA were 1-76±0-42, 1-83±0-55, 1-85±0-62, 7-36±0-92, 5-35±0-77, 5-06±0-62 and 821-68±95-12, 868-15±102-82, 723-39±92-56, 1 660-08±113-65, 1 433-01±107-24, 1 562-36±112-88 respectively- The increased concentrations of mitochondrial ROS and IL-1β upregulation were confirmed in the inhibition but not the induction of auto-phagy-We also found that downregulation of ATG5 and Beclin1 sensitized cells for the release of IL-1β induced by MSU (monosodium urate) or nigericin which was the NLRP3 inflammasome activator- RSV treatment attentuated this effect- Conclusion: Long-term application of high glucose-based peritoneal dialysis fluids can trigger the consistent activation of NLRP3-IL-1β in peritoneal mesothelial cells-Timely initiation of autophagy may block the NLRP3-IL-1β activation and provide a basis for the further development of a potential therapeutic strategy for delay of chronic inflammation and peritoneal fibrosis associated with peritoneal dialysis.