血管紧张素-(1-7)对人脐静脉平滑肌细胞钙化的影响及其机制研究  

Effects of Angiotensin-(1-7) on Vascular Calcification in Human Umbilical Vein Smooth Muscle Cells

作　　者：张娟[1] 张鸣号[1] 李慧[1] 李桂忠[1] 王秀梅[1] 曹军[1] 

机构地区：[1]宁夏医科大学基础医学院病理生理学系,银川750004

出　　处：《宁夏医科大学学报》2011年第12期1122-1126,F0002,共6页Journal of Ningxia Medical University

基　　金：宁夏高等学校科学技术研究项目

摘　　要：目的复制人脐静脉平滑肌细胞(HUSMCs)钙化模型,观察血管紧张素-(1-7)[Ang-(1-7)]对HUSMCs钙沉积的影响,探索Ang-(1-7)延缓或抑制HUSMCs钙化的机制。方法采用β-甘油磷酸盐(β-GP)复制HUSMCs钙沉积模型,Ang-(1-7)干预后,对细胞进行Von Kossa染色及图像分析、检测碱性磷酸酶(ALP)活性和细胞钙含量;检测细胞培养液中转化生长因子β1(TGFβ1)活性、骨钙素(OC)含量及细胞中核心结合因子1(Cbfα1)蛋白表达。结果 Ang-(1-7)可明显减轻钙化HUSMCs聚集生长,Von Kossa染色显示细胞聚集处棕褐色钙结节较钙化组明显减轻(P<0.01),且10-5、10-7、10-9mol.L-1Ang-(1-7)与钙化组相比钙化细胞百分比、细胞钙含量和ALP活性均明显降低(P均<0.01),细胞TGFβ1分泌水平、Cbfα1表达、骨钙素含量均较钙化组降低(P均<0.01)。结论 Ang-(1-7)可减轻HUSMCs钙化程度,其作用机制与减弱TGFβ1-Smads-Cbfα1信号通路中关键信号分子TGFβ1、Cbfα1的表达有关。Objective To observe the effect of Angiotensin-（1-7）[Ang-（1-7）] in vascular calcification and to explore the mechanism that Ang-（1-7） delay or inhibit the HUSMCs calcium deposition.Methods β-glycerolphosphate（β-GP） was used to stimulate the human umbilical vein cells which cultured in vitro.The cells were randomly divided into 6 groups（n=6）.Calcium deposition in HUSMCs were detected by Von Kossa staining and Von Kossa staining was analyzed by micro-image analysis system.The activities of alkaline phosphatase（ALP） in HUSMCs were detected by pheny1 diphosphate-2-sodium.The total calcium contents were calculated by ultraviolet spectrophotometry.Activity of TGFβ1 in the nutrient solution was monitored by ELISA.The expression of core binding factorα1（Cbfα1） in HUSMCs were analyzed by Westernblot.Osteocalcin（OC） contents in the nutrient solution were determined by radio-immunoassay.Results HUSMCs calcification were established successsfully.After Von Kossa staining,a large number of daisy-shaped HUSMCs were found in 12mmol·L-1 β-glycerolphosphate group.There were brown nodules among smooth muscle cells.Ang-（1-7） significantly reduced the calcification of the HUS-MCs.Compared with CAL group,Von Kossa staining of 10-5mol·L-1 Ang-（1-7） group,10-7mol·L-1 Ang-（1-7） group,brown nodules in 10-9mol·L-1 Ang-（1-7） group significantly reduced（respectively P〈0.01）;Compared with CAL group,the percentage of positive cells in calcification,the calcium content and ALP levels of 10-5mol·L-1 Ang-（1-7） group,10-7mol·L-1 Ang-（1-7） group,brown nodules in 10-9mol·L-1 Ang-（1-7） group significantly reduced（respectively P〈0.01）.Compared with the CAL group,TGFβ1 levels,expression of cbfα1 in HUSMCs and osteocalcin contents in culture medium of 10-5mol·L-1 Ang-（1-7） group,10-7mol·L-1 Ang-（1-7） group,10-9mol·L-1 Ang-（1-7） group significantly reduced respectively（respectively P〈0.01）.Conclusion Ang-（1-7） could inhibit the HUSMCs calcific

关 键 词：血管紧张素-(1-7) 血管钙化 转化生长因子Β1 核心结合因子1 骨钙素 

分 类 号：R363[医药卫生—病理学]