胰高血糖素样肽-1可改善高糖环境下人脐静脉内皮细胞的氧化应激

The Mitigation of GLP-1 on High Glucose-induced Human Umbilical Vein Endothelial Cell Oxidative Stress

目的探讨胰高血糖素样肽-1(GLP-1)对高糖培养的原代人脐静脉内皮细胞(HUVECs)氧化应激情况的影响。方法体外分离、培养原代人HUVECs,分为对照组、高糖组(30 mmol/L)及GLP-1预处理组(GLP-1 100nmol/L预处理1h后30mmol/L高糖培养)。检测细胞上清乳酸脱氢酶(LDH)漏出量,进行细胞毒性作用定量分析;化学发光法检测细胞内活性氧族(ROS)水平、黄嘌呤氧化酶法检测超氧化物歧化酶(SOD)活力了解细胞内氧化应激状态。结果 (1)GLP-1可以改善高糖环境对人脐静脉内皮细胞的毒性作用,高糖组LDH漏出量为对照组的1.8倍,GLP-1预处理组的LDH漏出量为对照组的1.3倍(P<0.05)。(2)高糖组细胞内ROS荧光强度较对照组增强1倍以上,而GLP-1预处理组ROS荧光强度可抑制至对照组水平(P<0.05);与对照组比较,高糖组细胞内SOD酶活性无明显改变;但与高糖组比较,GLP-1预处理组SOD酶活性升高了64.64%(P<0.05)。结论 GLP-1可以改善高糖对HUVECs的细胞毒性作用,其机制可能是通过减轻细胞氧化应激来实现的。

Objective To investigate the effects and mechanism of glucagon-like peptide-1 （GLP-1） on high glucose-induced human umbilical vein endothelial cells （HUVECs） oxidative stress . Methods HUVECs was isolated and cultured in vitro ,randomly divided into control group ,high glucose （30 mmol/L） group and GLP-1 group （100 nmol/L GLP-1＋ high glucose） . Endothelial toxicity was evaluated by lactate dehydrogenase （LDH） method . Intracellular reactive oxygen species （ROS） produc-tion and activity of superoxide dismutase （SOD） reflects the oxidative stress state in the cell . Fluores-cence enzyme-labeled instrument was used to detect the production of ROS . The activity of SOD was de-tected by xanthine oxidase method . Results （1）GLP-1 could reduce toxicity of high glucose . After high glucose trestment ,the ratio of LDH leakage was markly increased to 1 .8 times of the control group . However ,the ratio of LDH leakage in cells pre-treated with 100 nmol/L GLP-1 significantly decreased to 1 .3 times of the control group（P〈0 .05） . （2）Comparing with control group ,ROS production was in-creased more than 1 times（P〈0 .05） and the activity of SOD had no significant changes in high glucose group . After GLP-1 pretreatment ,ROS was lowered to the level of control group and activity of SOD was increased by 64 .64% （P〈 0 .05） . Conclusions GLP-1 has a direct protective effect on HUVECs function under high glucose condition ,w hich is probably related to the mitigation of oxidative stress .