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目的观察低浓度葡萄糖对人脐静脉内皮细胞一氧化氮(NO)与活性氧(ROS)变化的影响。方法以体外培养人脐静脉内皮细胞株HUVEC-12为研究对象,将实验分为正常对照组(5.5mmol/L葡萄糖)、低糖组(2.8mmol/L葡萄糖)、无糖组(0mmol/L葡萄糖)。分别用2.8、0mmol/L葡萄糖干预HUVEC-12细胞,经过1、2、4、12h后,硝酸还原酶法测定NO产量,化学比色法测定一氧化氮合成酶(NOS)活性,Dihydroethidium荧光探针法测定细胞内ROS水平。结果与正常对照组相比,低糖组与无糖组NO水平降低(P<0.01),NOS活性下降(P<0.01),细胞内ROS水平升高(P<0.01),均呈剂量依赖关系。同一浓度组内随着时间的延长,内皮细胞NO水平、NOS活性进一步降低(P<0.01),ROS水平进一步升高(P<0.05),各指标都具有浓度和时间依赖性。结论低糖可导致内皮细胞功能障碍,NO水平降低,NOS活性下降,其机制可能与低糖导致内皮细胞氧化应激损伤有关。
Objective To observe the effects of low concentration glucose on the changes of nitric oxide (NO) and reactive oxygen species (ROS) in human umbilical vein endothelial cells. Methods Human umbilical vein endothelial cell line HUVEC-12 was cultured in vitro. The experiment was divided into normal control group (5.5mmol / L glucose), low glucose group (2.8mmol / L glucose), glucose-free group ). HUVEC-12 cells were treated with glucose of 2.8 and 0mmol / L, respectively. After 1, 2, 4 and 12 h, NO production was measured by nitrate reductase method. Nitric oxide synthase (NOS) Acupuncture determination of intracellular ROS levels. Results Compared with the normal control group, the NO level decreased (P <0.01), the NOS activity decreased (P <0.01) and the intracellular ROS level increased (P <0.01) in a dose-dependent manner. With time prolonging in the same concentration group, the level of NO and the activity of NOS decreased (P <0.01) and the level of ROS increased further (P <0.05) in endothelial cells. Both indexes were concentration-dependent and time-dependent. Conclusion Low glucose can cause endothelial dysfunction, decrease the level of NO and decrease the activity of NOS. The mechanism may be related to the oxidative stress injury induced by low glucose in endothelial cells.