Yahyapour N, Eriksson C, Kjellstrand P, Wieslander A, Nygren H
Applied Cell Biology, Institute of Anatomy and Cell Biology, University of Göteborg, Sweden.
Toxicol In Vitro. 2001 Dec;15(6):655-61. doi: 10.1016/s0887-2333(01)00073-x.
The possibility of reducing the cytotoxic effect of heat-sterilized peritoneal dialysis (PD) fluid by addition of antioxidants/scavengers during incubation of titanium-adhering cells was investigated. Capillary blood from healthy donors was placed in drops on commercially available titanium pieces and incubated in a humidified chamber at 37 degrees C for 60min. After incubation the adherent polymorphonuclear leukocytes were immersed for 1-4h in PD-fluid, pH 7.4, containing 2.5% glucose with glutathione (GSH), superoxide dismutase, catalase or dithiothreitol (DTT). Luminol- or isoluminol-amplified chemiluminescence was used to measure the zymosan- and phorbol myristate acetate (PMA)-stimulated respiratory burst activity, as an indicator of the cytotoxicity of the PD-fluids. Heat sterilized PD-fluid had inhibitory effect on zymosan-induced respiratory burst and impaired both the extracellular and intracellular PMA-induced respiratory burst. Addition of GSH to the PD-fluid resulted in reduction of cytotoxical effects on the zymosan-induced and extracellular PMA-induced respiratory burst. The intracellular respiratory burst was not affected. The present results show that GSH and DTT have the ability to protect polymorphonuclear leukocytes against the cytotoxic effects of the PD-fluid by keeping the cell membrane in a reduced state.
研究了在钛黏附细胞孵育过程中添加抗氧化剂/清除剂以降低热灭菌腹膜透析(PD)液细胞毒性作用的可能性。将健康供体的毛细血管血滴在市售钛片上,于37℃的湿润培养箱中孵育60分钟。孵育后,将黏附的多形核白细胞浸入pH值为7.4、含2.5%葡萄糖以及谷胱甘肽(GSH)、超氧化物歧化酶、过氧化氢酶或二硫苏糖醇(DTT)的PD液中1 - 4小时。采用鲁米诺或异鲁米诺增强化学发光法测量酵母聚糖和佛波酯肉豆蔻酸酯(PMA)刺激的呼吸爆发活性,作为PD液细胞毒性的指标。热灭菌的PD液对酵母聚糖诱导的呼吸爆发有抑制作用,并损害细胞外和细胞内PMA诱导的呼吸爆发。向PD液中添加GSH可降低对酵母聚糖诱导的以及细胞外PMA诱导的呼吸爆发的细胞毒性作用。细胞内呼吸爆发未受影响。目前的结果表明,GSH和DTT能够通过使细胞膜保持还原状态来保护多形核白细胞免受PD液的细胞毒性作用。