Kuo Hung-Tien, Chen Hsiang-Wen, Hsiao Hui-Hsu, Chen Hung-Chun
Division of Nephrology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Faculty of Renal Care, Kaohsiung Medical University, Kaohsiung, Taiwan.
Nephrol Dial Transplant. 2009 Jun;24(6):1799-809. doi: 10.1093/ndt/gfn718. Epub 2009 Jan 6.
Chronic peritoneal dialysis (PD) is one of the major therapies for uremic patients. However, the peritoneal mesothelial cells (PMCs) are subject to the injury by bioincompatible dialysates. The aim of this study is to investigate the protective roles and mechanisms of heat shock response in PMCs.
Primary cultured human PMCs (HPMCs) were subjected to commercial peritoneal dialysates. The cell viability was assayed by MTT test and Annexin V assay. The expression of HSPs was detected by Western blots analysis. Intracellular hydrogen peroxide and superoxide anion were detected using H(2)DCFDA and dHE probe, respectively, with flow cytometry. The mitochondrial membrane potential (DeltaPsim) of HPMCs was evaluated using JC1 probe with flow-cytometry.
Exposure of HPMCs to 1.5%, 2.5%, and 4.25% dextrose, and 7.5% icodextrin dialysates, respectively, for 60 min resulted in significantly accumulation of intracellular reactive oxygen species (ROS), DeltaPsim loss, and cell death in HPMCs. Amino acid dialysates exhibited no significant cytotoxicity. Adjusting the acidity in 1.5% dextrose and icodextrin dialysate significantly attenuated the dialysate-induced ROS generation and cell death in HPMCs. Heat pretreatment (41 degrees C, 30 minutes), which induced HSP 27 and 72 syntheses, significantly attenuated the dialysate-induced intracellular ROS accumulation, Dym loss, and cell death in HPMCs.
In conclusion, the acidic bioincompatible dialysates induce oxidative stress, DeltaPsim loss, and subsequent cell death in HPMCs. Amino acid dialysates is more biocompatible than glucose and icodextrin dialysates to HPMCs. Heat shock response protects HPMCs from the bioincompatible dialysates-induced cellular damage.
慢性腹膜透析（PD）是尿毒症患者的主要治疗方法之一。然而，腹膜间皮细胞（PMC）会受到生物不相容性透析液的损伤。本研究旨在探讨热休克反应在PMC中的保护作用及机制。
将原代培养的人PMC（HPMC）暴露于市售腹膜透析液中。通过MTT试验和Annexin V试验检测细胞活力。通过蛋白质印迹分析检测热休克蛋白（HSP）的表达。分别使用H(2)DCFDA和dHE探针，通过流式细胞术检测细胞内过氧化氢和超氧阴离子。使用JC1探针通过流式细胞术评估HPMC的线粒体膜电位（ΔΨm）。
将HPMC分别暴露于1.5%、2.5%和4.25%的葡萄糖透析液以及7.5%的艾考糊精透析液中60分钟，导致HPMC内活性氧（ROS）显著积累、ΔΨm丧失和细胞死亡。氨基酸透析液未表现出明显的细胞毒性。调节1.5%葡萄糖和艾考糊精透析液的酸度可显著减轻透析液诱导的HPMC中ROS生成和细胞死亡。热预处理（41℃，30分钟）诱导HSP 27和72合成，显著减轻透析液诱导的HPMC内ROS积累、ΔΨm丧失和细胞死亡。
总之，酸性生物不相容性透析液在HPMC中诱导氧化应激、ΔΨm丧失及随后的细胞死亡。氨基酸透析液对HPMC的生物相容性高于葡萄糖和艾考糊精透析液。热休克反应可保护HPMC免受生物不相容性透析液诱导的细胞损伤。
