糖基化终产物对人脐静脉内皮细胞黏附分子表达和黏附功能的影响及其机制  

Effect and its mechanism of AGEs on expression of VCAM-1 and adhession of human vascular endothelial cells on monocytes

作　　者：刘云海[1] 徐标[1] 杨慎启[1] 蒲娟娟[1] 曹月新[1] 

机构地区：[1]东南大学心血管病研究中心

出　　处：《中国糖尿病杂志》2007年第4期239-242,共4页Chinese Journal of Diabetes

基　　金：国家自然科学基金资助项目(30170370)

摘　　要：目的探讨糖基化终产物(AGEs)对人脐静脉内皮细胞(HUVECs)血管细胞黏附分子1(VCAM-1)表达和黏附功能的影响及其机制。方法以D-葡萄糖和牛血清白蛋白在37℃条件下制备AGEs。胶原酶法分离HUVECs并加以培养。用流式细胞术测定HUVECs受AGEs干预后VCAM-1表达水平。按Esposito′s法进行HUVECs外周血单核细胞(PBMCs)黏附试验,Chen氏法计算HUVECs-PBMCs黏附率。用[γ-32P]ATP液闪计数技术测定受AGEs干预后HUVECsPKC活性的变化。结果AGEs可明显诱导HUVECsVCAM-1表达,呈剂量、时效依赖关系。HU-VECs-PBMCs黏附率的变化与VCAM-1的表达水平平行。蛋白激酶C(PKC)抑制剂可明显降低AGEs所致的VCAM-1表达增加及HUVECs-PBMCs黏附率增加。HUVECs受AGEs干预后其PKC活性明显升高。结论AGEs能使人血管内皮细胞VCAM-1表达增加和黏附功能增强,其机制是部分通过增加细胞内PKC的活性来实现。Objective To investigate effect and its mechanism of advanced glycosylation end products（AGEs） on expression of vascular cell adhesion molecule-1（VCAM-1） and activity of protein kinase C （PKC） in cultured human umbilical vein endothelial cells（HUVECs） as well as adhesion rate of HUVECs to human peripheral blood mononuclear cells（PBMCs）, and to explore influence of PKC inhibitor（STS） on above changes. Methods AGEs were made by incubating bovine serum albumin （BSA） with high concentration of glucose in 37℃ in vitro. HUVECs were cultured according to the descriptions in literatures. The production of VCAM-1 induced by AGEs was determined by flowcytometry（FCM）. Adhesion assay and adhesion rate of HUVECs on PBMCs were performed by Esposito＇s and Chen＇s method. The activity of PKC was detected through [γ-32P] ATP liquid scintillation counting. Results AGEs promoted HUVECs adhesion to PBMCs through upregulation expression of VCAM-1 in HUVECs. STS could markedly depress VCAM-1 expression and adhesion of HUVECs-PBMCs. The activity of PKC could be enhanced by AGEs. Conclusions AGEs can upregulate VCAM-1 expression and PKC activity of HUVECs, and increase adhesion of HUVECs to PBMCs. These effects can be incompletely blocked by STS. These suggest that AGEs increase VCAM-1 expression and adhesion rate of HUVECs to PBMCs partially through PKC pathway.

关 键 词：糖基化终产物 高级 脐静脉 内皮细胞 单核细胞 血管内皮细胞黏附分子-1 

分 类 号：R587.1[医药卫生—内分泌]