高糖诱导人脐静脉内皮细胞内皮微粒释放并促进凋亡的实验研究.doc

高糖诱导人脐静脉内皮细胞内皮微粒释放并促进细胞凋亡 田刚 任燕 卢群 鲁敏 西安交通大学医学院第一附属医院心内科 陕西 西安710061 摘要 目的：探讨高糖对人脐静脉内皮细胞(human umbilical vein endothelial cell , HUVEC)内皮微粒释放和细胞凋亡的影响。方法：用不同浓度的葡萄糖（5.5 mmol/L、 16.7 mmol/L和33.3 mmol/L）分别作用于培养成功的HUVEC 0 h、12 h、24 h、48 h和72 h。设立正常组（5.5 mmol/L葡萄糖）和甘露醇组（27.8 mmol/L）作为对照。应用MTT比色法检测细胞的生存率。采用流式细胞仪分别检测HUVEC内皮微粒释放以及细胞凋亡率。应用Griess还原法检测培养细胞上清液中一氧化氮（NO）浓度。结果：葡萄糖刺激诱导HUVEC内皮微粒的释放呈剂量和时间依赖性增高（P0.05）；不同浓度葡萄糖可不同程度促进HUVEC细胞凋亡,抑制HUVEC细胞增殖（P0.05）；高糖诱导的内皮微粒释放水平与细胞凋亡率呈正相关（r=0.659, P0.05），与NO浓度呈负相关（r=-0.873, P0.05）。结论：高葡萄糖刺激诱导人脐静脉内皮细胞内皮微粒释放，促进细胞凋亡。内皮微粒释放水平是反映内皮功能障碍及其程度的指标之一。 关 键 词：葡萄糖；糖尿病；动脉粥样硬化；内皮微粒；内皮功能 High Glucose Induced Endothelial Microparticles Released in Human Umbilical Vein Endothelial Cell and Promoted Cell Apoptosis TIAN Gang, REN Yan, LU Qun, LU Min Department of Cardiology，First Affiliated Hospital of Medical College，Xi’an Jiaotong University，Xi’an，Shaanxi，710061 [Abstract] Objects To explore the effects of high glucose on Endothelial Microparticles (EMPs) release and apoptosis in human umbilical vein endothelial cell (HUVEC). Methods The cells were incubated for 0 h，12 h，24 h，48 h and 72 h respectively with different concentrate of glucose (5.5 mmol/L，16.7 mmol/L and 33.3 mmol/L， respectively). The cells were cultured in the media containing glucose 5.5 mmol/L and mannitol 7.8 mmol/L as control groups. The survival rate of cells was examined by MTT assay. EMPs level and percent of cell apoptosis were detected by flow cytometry. Production of Nitric oxide (NO) in cell culture medium was determined by the Griess reaction technique. Results: The level of EMPs release induced by high glucose was significantly higher in concentration and time-dependent manner (P0.05). The different concentrate of glucose induced different cell apoptosis and inhibited cell proliferation(P0.05). It was a significant positive correlation among the EMPs released and cell apoptosis（r=0.659, P0.05）, and negative correlation between the EMPs released and NO level（r=-0.8