乳酸盐与丙酮酸盐腹膜透析液对外周血中性粒细胞凋亡的影响

Effects of glucose-based lactate and pyruvate peritoneal dialysates on peripheral blood neutrophils apoptosis

目的 :比较研究乳酸盐与丙酮酸盐腹膜透析 (腹透 )液对外周血中性粒细胞 (PMN)凋亡以及吞噬和杀菌能力的影响。方法 :含有不同浓度 (1.5 % ,4.2 5 % )葡萄糖的乳酸盐或丙酮酸盐腹透液 ,与外周血 PMN孵育不同时间 (10 ,30 m in)后 ,利用流式细胞术及荧光素标记的 Annexin- 检测细胞凋亡情况 ;通过菌落计数方法观察 PMN对金黄色葡萄球菌 (金葡菌 )吞噬和杀菌功能的改变 ,分别以细胞外和细胞内活菌下降的百分数表示。 结果 :PMN与 1.5 %乳酸盐腹透液 (p H6 .7)孵育 10min,收集细胞 ,培养 72 h后细胞凋亡率与正常对照组比较无差异 ,而孵育时间延长至 30 min后 ,培养的 PMN在 72 h凋亡率[(6 6 .90± 2 .35 ) % ]明显增加 ,与 4.2 5 %乳酸盐腹透液 (p H7.2 ) 10 m in组结果相似 [(6 5 .36± 2 .6 0 ) % ];1.5 %葡萄糖乳酸盐腹透孵育 30 m in及 4.2 5 %葡萄糖乳酸盐腹透液孵育 10 m in组 ,PMN的吞噬及杀菌功能均较对照组显著降低。无论是含1.5 %还是 4.2 5 %葡萄糖的丙酮酸盐腹透液 ,PMN凋亡率均与对照组无显著差异 ;对 PMN吞噬和杀金葡菌功能也无显著影响。 结论 :丙酮酸盐腹透液对外周血 PMN凋亡以及吞噬和杀菌功能的影响均较小 。

Objective: To compare the effects of variable glucose based lactate and pyruvate peritoneal dialysates on apoptosis, phagocytosis and bacteral killing of staphylococcus aureus in peripheral blood neutrophils. Methods: Cell apoptosis was analyzed by flow cytometry (FCM) to detect phosphatidylserine (PS) exposure on early apoptotic neutrophils using fluorescein labeled Annexin V. Phagocytosis and intracellular bactericidal assays were expressed with the decrease in the number of viable extracellular and intracellular bacteria. Results: After exposure to 1.5% glucose lactate dialysate (pH 6.7) for 10 min, neutrophils experienced spontaneously normal apoptosis. Prolonged to 30 min, however, neutrophils underwent accelerated apoptosis (66.90±2.35)% behaved like that exposure to 4.25% glucose lactate dialysate (pH 7.2) for 10 min (65.36± 2.60)% analyzed by FCM at 72 hours culture time. Nevertheless, for pyruvate based peritoneal dialysates, regardless of 1.5% or 4.25% glucose, there were little influences on neutrophils apoptosis. Lactate cytotoxicity of impaired phagocytosis and bacterial killing could be corrected by pyruvate based dialysates inspite of existence of hypertonic glucose. Conclusion: These data suggest that pyruvate based peritoneal dialysates have little effects on peripheral blood neutrophils either apoptosis or phagocytosis and bacterial killing. [