葡萄糖腹膜透析液对人腹膜间皮细胞线粒体活性氧的影响

The effect of mitochondrial reactive oxygen species production in human peritoneal mesothelial cells exposed to glucosebased peritoneal dialysis fluids

目的观察不同浓度葡萄糖腹膜透析液对人腹膜间皮细胞线粒体活性氧(ROS)的影响。方法体外培养人腹膜间皮细胞株第5至10代(HMr SV5,DMEM/F12培养基含10%胎牛血清)用于实验研究。MTT检测细胞活力,实验分为7组:对照组、1.5%Dextrose组、2.5%Dextrose组、4.25%Dextrose组、线粒体呼吸链复合酶Ⅰ抑制剂Rotenone组(5μM)、线粒体呼吸链复合酶Ⅱ抑制剂Thenoyltrifluoroacetone(TTFA)组(5μM)、线粒体呼吸链复合酶Ⅲ抑制剂Antimycin A组(100μg/ml),应用流式细胞仪检测总线粒体、呼吸线粒体、超氧化线粒体。结果各组作用12h,1.5%Dextrose、2.5%Dextrose、4.25%Dextrose、Rotenone、Antimycin A组总线粒体、超氧化线粒体明显增加,与对照组比较,差异有统计学意义(P<0.05),TTFA组与对照组比较无明显差异。结论葡萄糖腹膜透析液作用下人腹膜间皮细胞线粒体活性氧产生增加。

Objective To explore the effect of mitochondrial reactive oxygen species （ROS） production in human peritoneal mesothelial cells exposed to glucose-based peritoneal dialysis fluids. Methods HMrSV5 cells （SV40 immortalized hu- man peritoneal mesothelial cell line） were grown in type I collagen-coated dishes in DMEM/F12 containing 10% FCS. All ex- periments on HMrSV5 cells were performed between passage 5 and 10. Cell viability was assessed by MTT. Cells were divided into 7 groups： control group, 1.5%Dextrose group, 2.5%Dextrose group, 4.25%Dextrose group, Rotenone group, Thenoyltrifluo- roacetone （2＂TFA） group, Antimycin A group. Flow cytometric was used to analyze the respiring （Mitotracker deep red）, total （Mitotracker deep green） and ROS-generating mitochonclria （MitoSOX）. Results Treatment for 12h, total and ROS-generat- ing mitochondria were significantly increased in the cells exposed to 1.5%Dextrose group, 2.5%Dextrose group, 4.25%Dextrose group, Rotenone group and Antimyein A group when compared to the control group （P〈0.05）, and there was no obvious difference in TTFA group in contrast to control group. Conclusion Glucose-based peritoneal dialysis fluids can increase the mito- chondrial ROS production in human peritoneal mesothelial cells.