Overexpression of HSP-72 confers cytoprotection in experimental peritoneal dialysis
Overexpression of HSP-72 confers cytoprotection in experimental peritoneal dialysis
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DOI：
10.1111/j.1523-1755.2004.66040.x
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发表时间：
2004-12-01
影响因子：
19.6
通讯作者：
Aufricht, C
中科院分区：
文献类型：
--
作者：
Bidmon, B;Endemann, M;Aufricht, C
背景由于腹膜透析液的生物相容性低，腹膜透析会因间皮细胞损伤而变得复杂（PDF）。我们以前已经证明，热休克蛋白（HSP）-72是有力的上调，在腹膜透析的体外和体内模型中的间皮细胞PDF暴露的反应。本研究的目的是评估HSP-72过表达的潜在细胞保护作用。通过比较经过延长的、通常致死的PDF暴露时间（120 min，CAPD 2; Fresenius，Bad洪堡，德国）的预处理与未预处理的人间皮细胞（Met 5a; ATCC，Manassas，VA，USA和原代细胞培养物）之间的细胞活力来评估细胞保护作用。预处理通过暴露于PDF（60分钟，CAPD 2; Fresenius）或加热（15分钟，41.5 ℃）进行，并通过用HSP-72瞬时转染进行。当间皮细胞经PDF或热非致死性暴露预处理后，HSP-72表达显著上调（>5倍，P < 0.01）。预处理的人间皮细胞被显著保护免于随后的“致死”暴露于PDF，如通过染料排除（>50%减少，P < 0.05）和乳酸脱氢酶（LDH）释放（>30%减少，P < 0.05）评估的。用HSP-72瞬时转染后，培养的人间皮细胞中HSP-72的过表达（>5倍）表明了相当的细胞保护作用（通过染料排除减少50%）。HSP- 72和ApopTag（凋亡检测试剂盒）双染技术证实了这种细胞保护作用。因此，我们的研究表明，间皮应激反应赋予细胞保护实验性腹膜透析，介导的HSP- 72的诱导，和刺激的预处理不一定是相同的后续损伤。这些数据提供了一个有吸引力的新的治疗方法对PDF毒性的基础。
Background. Peritoneal dialysis is complicated by mesothelial cell injury due to low biocompatibility of peritoneal dialysis fluid (PDF). We have previously demonstrated that heat shock protein (HSP)-72 is potently up-regulated in response to PDF exposure of mesothelial cells in in vitro and in vivo models of peritoneal dialysis. The aim of this study was to evaluate potential cytoprotective effects of overexpression of HSP-72.Methods. Cytoprotection was assessed by comparing cellular viability between pretreated versus nonpretreated human mesothelial cells (Met 5a; ATCC, Manassas, VA, USA, and primary cell cultures) subjected to extended, usually lethal PDF exposure times (120 min, CAPD2; Fresenius, Bad Homburg, Germany). Pretreatment was performed with exposure to PDF (60 min, CAPD2; Fresenius) or heat (15 min, 41.5degreesC), and by transient transfection with HSP-72.Results. When mesothelial cells were pretreated by nonlethal exposure to PDF or heat, HSP-72 was markedly up-regulated (>5-fold, P < 0.01). Pretreated human mesothelial cells were significantly protected against subsequent "lethal" exposures to PDF, as assessed by dye exclusion (>50% reduction, P < 0.05) and lactate dehydrogenase (LDH) release (>30% reduction, P < 0.05). Comparable cytoprotection (50% reduction by dye exclusion) was indicated by overexpression of HSP-72 in cultered human mesothelial cells (>5-fold) after transient transfection with HSP-72. This cytoprotection was confirmed at a cellular basis by double staining techniques with HSP- 72 and ApopTag (apoptosis detection kit).Conclusion. Our study therefore shows that the mesothelial stress response confers cytoprotection in experimental peritoneal dialysis, mediated by the induction of HSP- 72, and that the stimulus of the pretreatment does not have to be identical to the subsequent injury. These data offer the basis for an attractive novel therapeutic approach against PDF toxicity.