D-葡萄糖对血管内皮细胞形态及其LOX-1蛋白表达的影响

摘要

目的：观察D-葡萄糖对血管内皮细胞形态及其血凝素样氧化低密度脂蛋白受体1（ LOX-1）表达的影响。方法取15、30、60 mmol／L D-葡萄糖（观察组）及等体积培养液（对照组）作用于人脐静脉内皮细胞（ HUVEC-12），采用倒置相差显微镜观察血管内皮细胞形态，Western blot 法检测细胞LOX-1蛋白相对表达。结果对照组细胞生长状态良好，多呈多角形或扁平形。细胞之间连接紧密，折光性强。观察组细胞排列较为紊乱，细胞核模糊，细胞间隙明显增大，折光性变弱，细胞内颗粒状物增多；其中以30 mmol／L D-葡萄糖作用48 h的细胞形态变化最明显。对照组与加入15、30、60 mmol／L D-葡萄糖的观察组作用48 h后，细胞LOX-1蛋白相对表达量分别为1．0、1．5±0．1、2．0±0．2、1．8±0．2；两两比较，P均＜0．05。观察组加入30 mmol／L D-葡萄糖分别作用0、12、24、48、72 h后LOX-1蛋白相对表达量分别为1．0、1．1±0．1、1．4±0．2、2．0±0．2、1．5±0．1；两两比较，P均＜0．05。结论 D-葡萄糖可导致血管内皮细胞损伤，同时上调细胞LOX-1蛋白表达。%Objective To observe the effects of D-glucose on the cell morphology and lectin-like oxidized low density lipoprotein receptor-1(LOX-1) expression of vascular endothelial cells .Methods Human umbilical vein endothelial cell line (HUVEC-12) was treated by D-glucose with different concentrations (15, 30 and 60 mmol/L) as the observation group, and the same amount of culture medium was used in the control group .The morphologic changes of the HUVEC-12 were observed under inverted phase contrast microscope and the expression of LOX -1 protein of them was detected by West-ern blotting .Results HUVEC-12 grew well in the control group and most of them were polygons or flattened .There were tight junction between endothelial cells and strong light refraction on the surface of them .While HUVEC-12 were less order-ly , the reflected light of cells tended to be weaker , the nuclei of them tended to be not clearly visible and the space between cells and particulate matter in cells tended to increase in the observation group .The morphology changes were the most ob-vious in the HUVEC-12 after being treated by 30 mmol/L D-glucose for 48 h.After being treated for 48 h, the LOX-1 pro-tein expression in the control group and observation group (15, 30 and 60 mmol/L) was 1.0, 1.5 ±0.1, 2.0 ±0.2 and 1.8 ±0.2 (all P<0.05).The relative LOX-1 protein expression of HUVEC-12 which were treated by 30 mmol/L D-glu-cose in the observation group for 0, 12, 24, 48 and 72 h were 1.0, 1.1 ±0.1, 1.4 ±0.2, 2.0 ±0.2 and 1.5 ±0.1(all P<0.05).Conclusion D-glucose can induce the injuries of vascular endothelial cells and up-regulate the expression of LOX-1.