心肌纤维化细胞模型的构建及缬沙坦对其治疗的研究  

Construction of Myocardial Fibrosis Cell Model and Study the Therapeutic Effect of Valsartan

作　　者：姜美华[1] 涂玲[1] 徐西振[1] 赵刚[1] 吴蕊[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院综合科,武汉430030

出　　处：《医药导报》2008年第7期739-742,共4页Herald of Medicine

基　　金：国家自然科学基金资助项目(基金编号:30470712)

摘　　要：目的构建心肌纤维化细胞模型,并探讨缬沙坦对其的作用。方法胶原酶消化法分离心肌成纤维细胞(cardiac fibroblasts,CFs),取第3或4代CFs加入10-6μmol·L-1血管紧张肽Ⅱ(angiotensinII,AngⅡ)构建心肌纤维化细胞模型。0.01,0.10,1.00,10.00,100.00μmol·L-1缬沙坦干预模型4,6,8,12,24h后光镜下观察细胞生长状况。采用四甲基偶氮唑盐(MTT)法及吖啶橙/溴化乙锭(AO/EB)染色法定量分析细胞活力,电镜技术观察亚细胞结构。结果CFs在种植3h后开始贴壁生长。加入AngⅡ后,光镜下可见细胞增生活跃。加入不同浓度的缬沙坦24h内,部分细胞呈凋亡的形态学改变。MTT提示缬沙坦浓度为10.00μmol·L-1作用8h后,CFs的生长抑制率最高。AO/EB染色表明上述条件下CFs死亡率最高,死亡方式主要为凋亡。电镜亦显示死亡细胞主要呈凋亡的形态学改变。结论在现有技术条件下体外构建心肌纤维化细胞模型完全可行。缬沙坦有效抑制模型中CFs增殖可能通过诱导其凋亡实现。Objective To construct the cell model of myocardial fibrosis （ MF）, investigate the therapeutic effect of valsartan on which, and provide a foundation for further research. Methods Cardiac fibroblasts （ CFs ） were isolated by collagenase digesting. The MF cell model was constructed with the addition of angiotensin Ⅱ （ Ang Ⅱ ） into the cells at the 3rd or 4th generation with a final concentration of 10^-6 μmol · L^-1. The growth condition of CFs was observed and recorded under the light microscope at 4,6,8,12 and 24 h after drug intervention. The viability of CFs was quantitatively assessed by MTT assay and AO/EB staining. Transmission electron microscopy was used to observe the subcellular structure of cells. Results The CFs began to adhere to the bottom 3 hours after plantation. The growth rate of the CFs was increased significantly with Ang 11 . Morphological alteration like apoptosis was showed in part of cells within 24 hours of Valsartan′s addition. The most powerful inhibition on CFs was determined by valsartan at a concentration of 10.00μmol · L^-1 after 8 hours of treatment by MTT assay. It was also suggested that CFs showed the highest mortality rate under this condition by AO/EB staining method, and the main cause of death was apoptosis. The results drawn form transmission electron microscopy also indicated that apoptosis was the major feature of cell death. Conclusion The cell model of MF could be constructed successfully under the available condition. Moreover, the CFs proliferation could be inhibited by valsartan for inducing the cell apoptosis.

关 键 词：缬沙坦 心肌纤维化 血管紧张肽Ⅱ 

分 类 号：R972.4[医药卫生—药品] R965.1[医药卫生—药学]