Marzec Lukasz, Liberek Tomasz, Chmielewski Michal, Bryl Ewa, Witkowski Jacek M, Liberek Krzysztof, Zdrojewski Zbigniew, Rutkowski Boleslaw
Department of Nephrology, Transplantology and Internal Medicine, and Department of Pathophysiology, Medical University of Gdansk, Ul. Debinki 7, 80-211 Gdansk, Poland.
Perit Dial Int. 2007 May-Jun;27(3):288-95.
One of the main limitations of peritoneal dialysis (PD) is deterioration of functional and morphological characteristics of the peritoneum. This complication appears to be related to the low biocompatibility profile of PD fluids. Recently, induction of the heat shock protein (HSP) stress response was demonstrated in cultured human mesothelial cells exposed to PD fluid in vitro. We investigated whether expression of heat shock protein 72 (HSP-72) in peritoneal macrophages is induced upon exposure to PD fluid during continuous ambulatory PD.
Peritoneal leukocytes were isolated from 4-hour dwell dialysate; peripheral blood mononuclear cells (PBMC) and peripheral blood monocytes isolated from the same patients were used as a control. In separate experiments, PBMC from healthy individuals were exposed in vitro to different PD fluids or to culture media. Expression of HSP-72 was assessed by Western immunoblotting, flow cytometry, and reverse-transcription polymerase chain reaction analysis.
Macrophages and leukocytes isolated from dialysis effluent expressed significantly increased HSP-72 and mRNA levels compared to blood monocytes and PBMC of the same patients. In vitro exposure of PBMC to fresh PD fluids resulted in significantly higher expression of HSP-72 compared to those incubated in culture medium. PBMC exposed in vitro to standard lactate-buffered dialysis fluids also expressed significantly more HSP-72 compared to cells exposed to bicarbonate/lactate-buffered fluids.
Our results indicate that exposure to PD fluids during dialysis triggers a shock response in peritoneal cells, which is manifested by significantly increased HSP-72 expression at both protein and mRNA levels. Analysis of this protein expression in peritoneal macrophages could be a new, convenient, and relevant way to assess the biocompatibility of PD fluids ex vivo.
腹膜透析（PD）的主要局限性之一是腹膜功能和形态特征的恶化。这种并发症似乎与腹膜透析液的低生物相容性有关。最近，在体外暴露于腹膜透析液的培养人腹膜间皮细胞中证实了热休克蛋白（HSP）应激反应的诱导。我们研究了在持续非卧床腹膜透析期间，腹膜巨噬细胞暴露于腹膜透析液时是否会诱导热休克蛋白72（HSP-72）的表达。
从4小时留存的透析液中分离腹膜白细胞；从同一患者分离的外周血单核细胞（PBMC）和外周血单核细胞用作对照。在单独的实验中，将健康个体的PBMC在体外暴露于不同的腹膜透析液或培养基中。通过Western免疫印迹、流式细胞术和逆转录聚合酶链反应分析评估HSP-72的表达。
与同一患者的血液单核细胞和PBMC相比，从透析流出液中分离的巨噬细胞和白细胞表达的HSP-72和mRNA水平显著增加。与在培养基中孵育的PBMC相比，将PBMC在体外暴露于新鲜腹膜透析液会导致HSP-72的表达显著更高。与暴露于碳酸氢盐/乳酸缓冲液的细胞相比，在体外暴露于标准乳酸缓冲透析液的PBMC也表达了显著更多的HSP-72。
我们的结果表明，透析期间暴露于腹膜透析液会触发腹膜细胞的应激反应，这表现为蛋白质和mRNA水平上HSP-72表达的显著增加。分析腹膜巨噬细胞中的这种蛋白质表达可能是一种新的、方便且相关的体外评估腹膜透析液生物相容性的方法。
