【摘要】 目的:探讨体外持续高葡萄糖浓度和葡萄糖浓度波动条件下,激活健康成人外周血白细胞(WBCs),观察细胞内还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)水平及活性氧(ROS)产量,对比两种条件对WBCs呼吸爆发的影响。方法:以体外培养健康成人外周血WBCs为研究对象,将实验分为正常对照组(5.5mmol/LD-葡萄糖),持续高糖组(16.7mmol/LD-葡萄糖)与波动组(5.5～16.7mmol/LD-葡萄糖交替),所有组处理24h后以fMLP干预30min,激活WBCs。紫外分光光度计在340nm处测细胞裂解上清液吸光值,计算WBCs内NADPH含量;流式细胞仪检测WBCs产生ROS的水平。结果:与正常对照组相比,持续高糖和波动高糖均能使经fMLP刺激后WBCs的NADPH和ROS水平降低(P<0.05),且波动组作用更明显(P<0.05)。结论:持续性高葡萄糖浓度和波动性高葡萄糖浓度均能使健康成人外周血WBCs内NADPH水平及ROS产生能力下降,波动性高葡萄糖浓度效果更加明显,导致WBCs呼吸爆发功能受损。更多还原

【Abstract】 Objective To observe intracellular NADPH level and ROS production by activating human peripheral white blood cells(WBCs)under the conditions of continuing high glucose and fluctuating high glucose levels after stimulation and to compare the impact of the two conditions on respiratory burst in WBCs.Methods Isolated healthy WBCs were incubated in the medium containing normal glucose(5.5 mmol/L D-Glu),continuing high glucose(16.7 mmol/L D-Glu),or fluctuating high glucose(5.5 mmol/L and 16.7 mmol/L alternatively) for 24 h and then intervened with fMLP for 30 min for activation.Intracellular NADPH contents were assayed by measuring the absorbance of the cell supernatant at 340nm on spectrophotometer.Intracellular ROS was evaluated by flow cytometry.Results As compared with the normal control group,NADPH contents and ROS production in the cells were decreased in the group with continuing high glucose or with fluctuating high glucose after fMLP intervention(P<0.05) and the effect of fluctuating glucose was more obvious(P < 0.05).Conclusions Continuing high glucose and fluctuating high glucose concentration decrease intracellular NADPH contents and the ability of WBCs to produce ROS.The effect of the latter one was more obvious,leading to the impairment of respiratory burst in the cells.更多还原