Evidence for HSP-mediated cytoskeletal stabilization in mesothelial cells during acute experimental peritoneal dialysis
Evidence for HSP-mediated cytoskeletal stabilization in mesothelial cells during acute experimental peritoneal dialysis
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DOI：
10.1152/ajprenal.00503.2005
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发表时间：
2007-01-01
影响因子：
4.2
通讯作者：
Aufricht, Christoph
中科院分区：
文献类型：
--
作者：
Endemann, Michaela;Bergmeister, Helga;Aufricht, Christoph
腹膜透析液(PDF)生物相容性低会损伤间皮细胞并激活其应激反应。在这项研究中，我们研究了应激反应的主要细胞保护效应因子热休克蛋白(HSP)在实验性腹膜透析中间皮细胞骨架稳定中的作用。在培养的人间皮细胞中，通过体外PDF暴露后洗涤剂对标记蛋白的提取能力来评估细胞骨架的完整性。以细胞骨架蛋白片段与重组HSP-72或HSP-72抗体共孵育为基础，通过条件化(PDF预处理)和修复实验评价HSP对Ezrin的稳定作用。在大鼠模型中，在体内PDF暴露期间，间皮细胞从其腹膜单层中分离出来，在有和没有HSP-72过表达的情况下进行了评估(通过热调节)。在体外，通过显著改变Ezrin和ZO-1的洗涤剂提取能力，细胞骨架对亚致死性PDF暴露的破坏被证明。在恢复期间，HSP的恢复与显著的诱导和细胞骨架重新分布有关。条件化实验和体外修复实验均为HSP-72介导的细胞骨架稳定提供了证据。在大鼠模型中，热处理后HSP-72的过表达导致体内暴露于PDF的间皮细胞脱离显著减少。我们的结果证实了热休克蛋白在急性体外和体内暴露PDF后对间皮细胞细胞骨架完整性的修复和细胞保护中的重要作用。在临床环境中，反复暴露于PDF不仅可能对间皮细胞造成反复损伤，而且代表了一种无意的条件性治疗。
Low biocompatibility of peritoneal dialysis fluid (PDF) injures mesothelial cells and activates their stress response. In this study, we investigated the role of heat shock proteins (HSP), the main cytoprotective effectors of the stress response, in cytoskeletal stabilization of mesothelial cells in experimental peritoneal dialysis. In cultured human mesothelial cells, cytoskeletal integrity was assessed by detergent extractability of marker proteins following in vitro PDF exposure. Effects of HSP on stabilization of ezrin were evaluated by a conditioning protocol (PDF pretreatment) and repair assay, based on coincubation of cytoskeletal protein fractions with recombinant HSP-72 or HSP-72 antibodies. In the rat model, detachment of mesothelial cells from their peritoneal monolayer during in vivo PDF exposure was assessed with and without overexpression of HSP-72 (by heat conditioning). In vitro, cytoskeletal disruption on sublethal PDF exposure was demonstrated by significantly altered detergent extractability of ezrin and ZO-1. Restoration was associated with significant induction and cytoskeletal redistribution of HSP during recovery. Both the conditioning protocol and in vitro repair assay provided evidence for HSP-72-mediated cytoskeletal stabilization. In the rat model, overexpression of HSP-72 following heat conditioning resulted in significantly reduced detachment of mesothelial cells on in vivo exposure to PDF. Our results establish an essential role of HSP in repair and cytoprotection of cytoskeletal integrity in mesothelial cells following acute in vitro and in vivo exposure to PDF. Repeated exposure to PDF, as is the rule in the clinical setting, may not only cause repeat injury to mesothelial cells but rather represents a kind of inadvertent conditioning treatment.