【作者】 桑丹； 薛耀明； 沙建平； 龙可； 付霞军； 李际敏；

【Author】 SANG Dan;XUE Yao-ming;SHA Jian-ping;Zhou Lin;Gao Fang;Long Ke;FU Xia-jun;Department of Endocrinology and Metabolism,Nanfang Hospital,Southern Medical University;

【机构】 南方医科大学南方医院内分泌代谢科；

【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,激活健康人外周血白细胞(WBCs),观察细胞内葡萄糖-6-磷酸脱氢酶(G6PD)活性、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,对比两种条件对白细胞磷酸戊糖途径及呼吸爆发的影响。方法:以体外培养健康人外周血WBCs为研究对象,将实验分为正常对照组(5.5mmol/L D-葡萄糖),持续高糖组(16.7mmol/L D-葡萄糖)与波动组(5.5-16.7 mmol/L D-葡萄糖交替),所有组处理24h后以fMLP干预30min,激活WBCs。四氮唑蓝定量测定法测定G6PD活性;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WBCs的G6PD活性、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人外周血WBCs内G6PD活性、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 investigate the effects of continuing high glucose and fluctuating high glucose on glucose-6-phosphate dehydrogenase(G6PD) activity、reduced form of nicotinamide-adenine dinucleotide phosphate(NADPH) level and the production of reactive oxygen species(ROS) of healthy human peripheral white blood cells(WBCs) after stimulation and respiratory burst ability of WBCs. Methods: Isolated healthy human peripheral WBCs were incubated in the medium containing normal glucose(5.5mmol/L D-Glu),continuing high glucose(16.7mmol/L D-Glu) and fluctuating high glucose(5.5mmol/L and 16.7mmol/L alternative action) for 24 h with fMLP intervention for 30 min to stimulate healthy human peripheral WBCs.Intracellular G6 PD activity was measured by spectrophotometer.NADPH contents was assayed by measuring the absorbance of cell supernatant at 340 nm on spectrophotometer. Intracellular ROS was evaluated on flow cytometry. Results: Compared with the normal control group,D6 PD activity、NADPH contents and production of ROS of the cells were both found intervention(P<0.05) and fluctuating high glucose had more obvious effects(P<0.05). Conclusions Healthy human peripheral WBCs exposed to continuing high glucose and fluctuating high glucose concentration both have decreased D6 PD activity、NADPH contents and the ability of ROS production,the latter condition had more obvious effects leading to the impairment of respiratory burst of WBCs.更多还原