晚期糖基化终产物对人内皮细胞表达MCP-1和VCAM-1的影响及阿托伐他汀的干预  

The Effects of Advanced Glycation End Products on MCP-1 and VCAM-1 Expressions in Human Umbilical Vein Endothelial Cell and the Atorvastatin Intervention Effect

作　　者：王科峰[1] 徐尚华[1] 许昌声 谢良地 

机构地区：[1]福建医科大学附属南平第一医院心内科,福建省南平市353000 [2]福建省高血压研究所,福建省福州市350004

出　　处：《中国动脉硬化杂志》2011年第4期299-304,共6页Chinese Journal of Arteriosclerosis

基　　金：福建省科技厅基金项目[(2007)3129]

摘　　要：目的观察晚期糖基化终产物对体外培养人脐静脉内皮细胞表达单核细胞趋化蛋白1和血管细胞黏附分子1的影响及阿托伐他汀的干预作用,探讨晚期糖基化终产物诱发动脉粥样硬化形成的机制和阿托伐他汀治疗的机制。方法胶原酶消化法分离获取人脐静脉内皮细胞;倒置相差显微镜形态观察法及免疫细胞化学染色法鉴定人脐静脉内皮细胞;逆转录聚合酶链反应法对单核细胞趋化蛋白1、血管细胞黏附分子1及晚期糖基化终产物受体基因表达进行分析;活性氧检测试剂盒检测细胞内活性氧产生量,倒置荧光显微镜下观察细胞内活性氧荧光强度。结果倒置相差显微镜下可见培养的细胞呈卵圆形或多角形,典型的铺路石样排列,经CD31、CD34染色,细胞浆内有棕黄色颗粒沉着（CD31、CD34阳性）。与空白对照组相比,晚期糖基化终产物（10^-4-10^-1g/L）呈浓度依赖性增强人内皮细胞单核细胞趋化蛋白1和血管细胞黏附分子1基因表达,10^-4g/L晚期糖基化终产物组人内皮细胞单核细胞趋化蛋1和血管细胞黏附分子1基因表达水平开始显著增高（0.26±0.02比0.17±0.04;0.22±0.02比0.08±0.01,P〈0.01）;与晚期糖基化终产物组相比,阿托伐他汀（0.1、1、10μmol/L）呈浓度依赖性抑制人内皮细胞单核细胞趋化蛋白1和血管细胞黏附分子1基因的表达,1μmol/L阿托伐他汀组人内皮细胞单核细胞趋化蛋白1和血管细胞黏附分子1基因表达水平开始显著降低（0.63±0.11比1.03±0.07;0.21±0.03比0.83±0.10,P〈0.01）;晚期糖基化终产物组人内皮细胞内活性氧荧光强度显著增强,阿托伐他汀干预后细胞内活性氧荧光强度明显降低,且0.1μmol/L阿托伐他汀组人内皮细胞晚期糖基化终产物受体基因表达水平显著降低（0.63±0.05比1.19±0.12,P〈0.01）。结论晚期糖基化终产物显著增强人内皮细胞单核细胞趋化蛋白1和血管细胞黏附分子1�Aim To investigate the effects of advanced glycation end products（AGE） on the expression of monocyte chemoattractant protein-1（MCP-1） and vascular cell adhesion molecule-1（VCAM-1） in human umbilical vein endothelial cell（HUVEC）and the intervention effect of atorvastatin. Methods Collagenase was used to isolate the endothelial cell from human umbilical vein;HUVEC were identified by immunocellochemistry and morphology;RT-PCR was performed to detect MCP-1,VCAM-1 and recepter for AGE（RAGE） mRNA expression;Reactive oxygen species（ROS） detectionkit was used to examine the level of ROS in HUVEC and inversion fluoescence microscope was used to observe the ROS level. Results The cultured cells were oval or polygon and retained cobblestone appearance through inverted phase contrast microscope,brown particles could be observed in cytoplasm（CD31 or CD34 positive cells） after immunocytochemical staining with CD31 or CD34;In comparison with control group,AGE（10^-4-10^-1 g/L）promoted MCP-1 and VCAM-1 mRNA expression in a concentration-dependent manner in HUVEC.The expression of MCP-1 and VCAM-1 mRNA was significantly elevated by AGE（10^-4 g/L） compared with the control group（0.26±0.02 vs 0.17±0.04;0.22±0.02 vs 0.08±0.01,P0.01）;Compared with AGE group,atorvastatin（0.1,1,10 μmol/L） diminished MCP-1 and VCAM-1 mRNA expression induced by AGE in HUVEC in a concentration-dependent manner;Atorvastatin in a dose of 1 μmol/L,could significantly decrease the expression of MCP-1 and VCAM-1 mRNA induced by AGE（0.63±0.11 vs 1.03±0.07;0.21±0.03 vs 0.83±0.10,P0.01）;The level of ROS in AGE group was higher than that in control group,atorvastatin could obviously decline the ROS level induced by AGE in HUVEC.Furthermore,atorvastatin（0.1 μmol/L） was able to decrease RAGE mRNA expression remarkly induced by AGE in HUVEC（0.63±0.05 vs 1.19±0.12,P0.01）. Conclusion AGE could significantly increase MCP-1 and VCAM-1 mRNA expression in HUVEC;Atorvastatin could decrease the oxidative str

关 键 词：动脉粥样硬化 晚期糖基化终产物 阿托伐他汀 晚期糖基化终产物受体 活性氧 单核细胞趋化蛋白1 血管细胞黏附分子1 

分 类 号：R541[医药卫生—心血管疾病]