[Effect of high glucose-based peritoneal dialysis fluids on NLRP3-IL-1β in human peritoneal mesothelial cells].

作者：X Y Li, Jun Wu, Dan Luo, W X Chen, Guyu Zhu, Y X Zhang, Zhao-yu Bi, Baohong Feng · 发表于：PubMed · 年份：2017 · 被引用次数：7 · 研究领域：Dialysis and Renal Disease Management、Kidney Stones and Urolithiasis Treatments、Inflammasome and immune disorders

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 com...