Department of Pediatrics and Adolescent Medicine, Medical University of Vienna, Währinger Gürtel 18-20, 1090, Vienna, Austria.
Cell Stress Chaperones. 2013 Sep;18(5):569-79. doi: 10.1007/s12192-013-0410-6. Epub 2013 Mar 14.
Non-physiological components of peritoneal dialysis fluids (PDF) lead to the injury of peritoneal mesothelial cells resulting in the failure of peritoneal dialysis (PD) potentially via inadequate induction of the protective heat shock response (HSR). Glycogen synthase kinase-3β (GSK-3β) is a negative regulator of cell survival partly by suppression of the HSR and is influenced by stress stimuli also present in conventional PDF. The effects of PDF on GSK-3β activation and the impact of GSK-3β inhibition with lithium (LiCl) were investigated on cell survival with special regard to HSR, in particular to heat shock transcription factor 1 (HSF-1) activation and Hsp72 production in an in vitro model of PD using MeT-5A and primary mesothelial cells. Incubation of cells with the PDF Dianeal® (glucose-based, low pH, high glucose degradation products (GDP)) and Extraneal® (icodextrin-based, low pH, low GDP) caused activation of GSK-3β compared to the other tested PDF, i.e. Balance®, Physioneal® (normal pH, glucose-based, low GDP) and Nutrineal® (moderately acidic, amino acid-based). Inhibition of GSK-3β with LiCl in Dianeal® and Extraneal®-treated cells dose-dependently decreased cell damage and death rate and was paralleled by higher HSF-1 activation and Hsp72 expression. GSK-3β is activated by low pH GDP containing PDF with and without glucose as osmotic agent, indicating that GSK-3β is involved in mesothelial cell signalling in response to experimental PD. Inhibition of GSK-3β with LiCl ameliorated cell injury and improved HSR upon PDF exposure. Thus, GSK-3β inhibitors likely have therapeutic potential as cytoprotective additive for decreasing PDF toxicity.
腹透液(PDF)中的非生理成分导致腹膜间皮细胞损伤,从而导致腹膜透析(PD)失败,这可能是由于保护性热休克反应(HSR)诱导不足所致。糖原合成酶激酶-3β(GSK-3β)是细胞存活的负调节剂,部分通过抑制 HSR 来实现,并且受到应激刺激的影响,这些应激刺激也存在于常规 PDF 中。本研究旨在探讨 PDF 对 GSK-3β激活的影响,以及用锂(LiCl)抑制 GSK-3β对细胞存活的影响,特别关注 HSR,特别是热休克转录因子 1(HSF-1)的激活和 Hsp72 的产生,采用 MeT-5A 和原代间皮细胞的 PD 体外模型进行研究。与其他测试的 PDF(即 Balance®、Physioneal®(正常 pH 值、基于葡萄糖、低 GDP)和 Nutrineal®(适度酸性、基于氨基酸)相比,细胞与 Dianeal®(基于葡萄糖、低 pH 值、高葡萄糖降解产物(GDP))和 Extraneal®(基于 icodextrin、低 pH 值、低 GDP)孵育会导致 GSK-3β 的激活。Dianeal®和 Extraneal®处理的细胞中,LiCl 抑制 GSK-3β 呈剂量依赖性降低细胞损伤和死亡率,并伴有更高的 HSF-1 激活和 Hsp72 表达。含有 GDP 的低 pH 值 PDF 可激活 GSK-3β,无论是否含有葡萄糖作为渗透剂,这表明 GSK-3β 参与了间皮细胞对实验性 PD 的信号转导。用 LiCl 抑制 GSK-3β 可减轻 PDF 暴露时的细胞损伤并改善 HSR。因此,GSK-3β 抑制剂作为减少 PDF 毒性的细胞保护添加剂具有治疗潜力。