胰高血糖素样肽-1改善过氧化氢诱导的血管内皮氧化损伤

发布时间：2018-04-22 12:30

本文选题：胰高血糖素样肽- + 过氧化氢　；参考：《暨南大学学报(自然科学与医学版)》2017年01期

【摘要】：目的:探讨胰高血糖素样肽-1(GLP-1)对过氧化氢(H2O2)诱导的内皮细胞氧化损伤的保护作用及机制.方法:取人脐静脉内皮细胞(HUVECs),分为正常对照组、H2O2组、GLP-1组、H2O2+浓度10 nmol/L GLP-1组、H2O2+浓度100 nmol/L GLP-1组、H2O2+浓度1 000 nmol/L GLP-1组.采用荧光显微镜检测细胞内活性氧自由基(ROS)水平、观察细胞骨架F-actin变化;采用流式细胞仪检测细胞凋亡比率;Western Blotting检测磷酸化的雷帕霉素靶蛋白(p-m TOR)蛋白水平;实时荧光定量聚合酶链反应(QPCR)检测抗氧化酶:过氧化氢酶(CAT)、超氧化物歧化酶2(SOD2)和谷胱甘肽过氧化物酶1(GPX1)基因的表达变化.结果:H2O2组细胞骨架破坏明显,细胞内ROS含量增加,细胞凋亡率增加;加入GLP-1后,细胞骨架破坏受到显著抑制,ROS含量减少,凋亡率降低.Western Blotting结果显示,H2O2组p-m TOR表达降低,GLP-1可以提高p-m TOR表达,使蛋白水平趋于正常;加入GLP-1受体拮抗剂艾塞纳肽(9~36)后,GLP-1不能上调p-m TOR表达.H2O2作用后,CAT、SOD2和GPX1基因表达降低,而GLP-1可以逆转这些基因表达的降低.结论:GLP-1通过上调抗氧化应激酶对抗H2O2诱导的内皮细胞损伤,且可能是通过活化m TOR通路起保护作用.
[Abstract]:Aim: to investigate the protective effect and mechanism of glucagon like peptide-1 (GLP-1) on oxidative injury of endothelial cells induced by hydrogen peroxide (H 2O 2). Methods: human umbilical vein endothelial cells (HUVECs) were divided into normal control group (H 2O 2 group), GLP-1 group (n = 10 nmol/L), H _ 2O _ 2 concentration (n = 10 nmol/L), H _ 2O _ 2 concentration (n = 100 nmol/L GLP-1), H _ 2O _ 2 concentration (n = 1 000 nmol/L GLP-1). The intracellular reactive oxygen free radical (Ros) level was detected by fluorescence microscopy, the cytoskeleton F-actin was observed by flow cytometry, and the phosphorylated rapamycin target protein was detected by flow cytometry (FCM) and phosphorylated rapamycin target protein (rapamycin). Real-time quantitative polymerase chain reaction (PCR) was used to detect the expression of antioxidant enzymes: catalase (catalase), superoxide dismutase (SOD) and glutathione peroxidase (Glutathione peroxidase 1 Glutathione). Results the cytoskeleton destruction was obvious, the ROS content was increased, the apoptosis rate was increased, and the cytoskeleton destruction was significantly inhibited by adding GLP-1. The results showed that the expression of p-m TOR in H _ 2O _ 2 group could increase the expression of p-m TOR and make the protein level normal, and the expression of p-m TOR and GPX1 could not be upregulated by adding GLP-1 receptor antagonist Eisenopeptide 936), and the expression of CATH-SOD2 and GPX1 was decreased after treatment with H _ 2O _ 2, the expression of p-m TOR could not be upregulated by the addition of GLP-1 receptor antagonist Eisenopeptide 936). GLP-1 can reverse the decrease in the expression of these genes. Conclusion 1: GLP-1 antagonizes endothelial cell injury induced by H2O2 by up-regulation of antioxidant responsive kinase, and may play a protective role by activating m TOR pathway.
【作者单位】：南方医科大学检验与生物技术学院生物治疗所;
【基金】：国家重大科学973研究计划项目(2011CB504006)
【分类号】：R587.1

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