摘要
背景:研究表明,糖化白蛋白在内皮细胞凋亡过程中发挥重要作用,并通过增加丝裂原活化蛋白激酶、酪氨酸激酶活性和产生活性氧产物介导单核细胞趋化因子1。目的:观察糖化白蛋白对培养的人脐静脉内皮细胞的单核细胞趋化因子1表达的影响。设计、时间及地点:单一样本观察,于2006-05/11在东南大学心血管实验室完成。材料:脐静脉内皮建株人脐静脉内皮细胞ECV304,引自中国科学院上海细胞生物研究所。方法:实验分两部分,第一部分以糖化白蛋白(浓度为400mg/L)对人脐静脉内皮细胞分别干预0,8,16,24,48,72h;第二部分以不同浓度(100,200,400,800mg/L)的糖化白蛋白对人脐静脉内皮细胞干预24h。两部分实验均用无血清培养基RPMI 1640和不含牛血清白蛋白葡萄糖干预(浓度为400mg/L)的无血清培养基作为对照。主要观察指标:四甲基偶氮唑盐微量酶反应比色法检测人脐静脉内皮细胞增殖,酶联免疫吸附法检测人脐静脉内皮细胞上清液单核细胞趋化因子1含量。结果:①不同时间点糖化白蛋白对人脐静脉内皮细胞增殖的抑制作用依次为48h>24h>16h(相临时间点比较,P均<0.05);干预8h对细胞的抑制无明显作用,72h抑制作用弱于48h。②与RPMI 1640组和牛血清白蛋白组相比,糖化白蛋白400mg/L及800mg/L对细胞的抑制作用明显增强(P<0.05),并随着糖化白蛋白浓度的增高而增强。③用浓度为400mg/L的糖化白蛋白干预6h及12h,人脐静脉内皮细胞单核细胞趋化因子1表达持续升高(P<0.05);随着糖化白蛋白浓度加大,细胞的单核细胞趋化因子1表达进一步增高,800mg/L时达到最高峰。结论:①糖化白蛋白以浓度、时间依赖方式抑制人脐静脉内皮细胞增殖,并在一定时间范围内呈时间依赖性地增加人脐静脉内皮细胞表达单核细胞趋化因子1表达。
BACKGROUND： Previous study reveals that, glycated albumin plays an important role on the apoptosis of endothelial cells and the expression of monocyte chemoattractam protein-1 through increasing the activity of mitogen activated protein kinase and protein tyrosine kinase, besides generating active oxygen products.
OBJECTIVE： To investigate the effects of glycated albumin on the expression of monocyte chemoattractant protein-1 in the cultured human umbilical vein endothelial cells （HUVECs）.
DESIGN, TIME AND SETTING： A single sample observation was carried out in the Cardiovascular Laboratory of Southeast University （Nanjing, Jiangsu, China） from May to November in 2006.
MATERIALS： ECV304 HUVEC strain was provided from Shanghai Institute of Cell Biology, Chinese Academy of Sciences （China）.
METHODS： Experimental procedures were assigned to two parts. On one hand, HUVECs were cultured with glycated albumin of the concentration of 400 mg/L for 0, 8, 16, 24, 48, or 72 hours. On the other hand, HUVECs were cultured with glycated albumin of the concentrations of 100, 200, 400, and 800 mg/L for 24 hours. In the control group, HUVECs were incubated with bovine serum albumin of the concentration of 400 mg/L and RPMI 1640 culture medium without addition of the serum.
MAIN OUTCOME MEASURES： The proliferation of HUVECs was estimated by MTT colorimetric assay. The content of monocyte chemoattractant protein-1 in the supernatant was measured by enzyme-linked immunosorbent assay.
RESULTS： Glycated albumin inhibited the proliferation rate of HUVECs in a time-dependent manner （P 〈 0.05）. The proliferation of cells cultured for 8 hours were not remarkably inhibited. However, the proliferation inhibiting effect on the cells cultured for 48 hours was significantly higher than that cultured for 72 hours. Compared to RPMI 1640 group and bovine serum albumin group, the proliferation inhibiting effect on cells cultured with 400 or 800 mg/L glycated albumin was significantly enhanced （P 〈 0.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第28期5422-5426,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
江苏省卫生厅开放课题基金(WK0510)~~
