Dobos G J, Andre M, Böhler J, Norgauer J, Lubrich-Birkner I, Steinhauer H B, Schollmeyer P J
Department of Nephrology, University Hospital Freiburg, Germany.
Adv Perit Dial. 1993;9:307-11.
Chemotaxis and phagocytosis are important functions of phagocytic cells, which are closely related to cytoskeletal reorganization. These functions may be abnormal in phagocytes of uremic patients undergoing continuous ambulatory peritoneal dialysis (CAPD). In order to examine whether these abnormalities result from treatment, we studied actin polymerization (AP), as an index of cytoskeletal alterations, chemotaxis, and phagocytosis in polymorphonuclear neutrophils (PMNs) of healthy subjects. Polymorphonuclear neutrophils were exposed to either a hepes buffer or a glucose-based dialysis solution (GBDS) of different pH's (5.2, 7.4) and different glucose concentrations (1.36%, 2.27%, 3.86%). After incubation for 0, 5, or 20 minutes, cells were activated with 10 nmol/L C5a-complement. AP was measured as filamentous (F) actin content by NBD phallacidin staining and FACS analysis. Chemotaxis of PMNs was measured in Boyden chambers. In addition, phagocytosis of zymosan particles was assessed. Prior exposure to GBDS pH 5.2 of each glucose concentration immediately and completely inhibited AP in response to 10 nmol/L C5a-complement, reduced chemotaxis (> 95%), and completely inhibited phagocytosis. The inhibition was pH-dependent, since GBDS pH 7.4 caused less inhibition of these functions. We conclude that glucose-based dialysis solutions are cytotoxic towards neutrophils and completely inhibit their ability to display responses requiring cytoskeletal reorganization.
趋化作用和吞噬作用是吞噬细胞的重要功能,它们与细胞骨架重组密切相关。在接受持续性非卧床腹膜透析(CAPD)的尿毒症患者的吞噬细胞中,这些功能可能会出现异常。为了研究这些异常是否由治疗引起,我们以健康受试者多形核中性粒细胞(PMN)中的肌动蛋白聚合(AP)作为细胞骨架改变的指标,对趋化作用和吞噬作用进行了研究。将多形核中性粒细胞暴露于不同pH值(5.2、7.4)和不同葡萄糖浓度(1.36%、2.27%、3.86%)的羟乙基哌嗪乙磺酸缓冲液或基于葡萄糖的透析液(GBDS)中。孵育0、5或20分钟后,用10 nmol/L C5a补体激活细胞。通过NBD鬼笔环肽染色和流式细胞术分析,将AP测定为丝状(F)肌动蛋白含量。在Boyden小室中测量PMN的趋化作用。此外,评估酵母聚糖颗粒的吞噬作用。预先暴露于每种葡萄糖浓度的pH 5.2的GBDS中,会立即并完全抑制对10 nmol/L C5a补体的反应中的AP,降低趋化作用(>95%),并完全抑制吞噬作用。这种抑制作用是pH依赖性的,因为pH 7.4的GBDS对这些功能的抑制作用较小。我们得出结论,基于葡萄糖的透析液对中性粒细胞具有细胞毒性,并完全抑制其表现出需要细胞骨架重组的反应的能力。