摘要
目的:建立血管紧张素Ⅱ(AngⅡ)诱导H9c2心肌细胞肥大模型,并观察各组心肌细胞泛素化结蛋白水平的变化。方法:常规培养H9c2心肌细胞,分别以3个AngⅡ浓度组(10^(-8)、10^(-7)、10^(-6) mol/L)分别处理不同时间(0、6、12、24、48、72 h),另设一个空白对照组;BCA法测量其总蛋白含量;并测量相同面积下心肌细胞数,计算心肌细胞相对表面积;Western blot法及免疫细胞化学法检测泛素化结蛋白的表达情况。结果:AngⅡ呈时间和浓度依赖关系地诱导H9c2心肌细胞肥大,表现为细胞总蛋白含量、细胞相对表面积和泛素化结蛋白表达均较对照组显著增多(P<0.05),且以10^(-7) mol/L作用48 h最为显著。结论:10^(-7)mol/L AngⅡ诱导H9c2心肌细胞48 h后成功建立心肌细胞肥大模型,且H9c2心肌细胞肥大后,泛素化结蛋白也明显升高。
Objective To establishment the model of angiotensin Ⅱ（AngⅡ） inducinged cardiomyocyte hypertrophy in H9c2 myocardial cells, and observinge the the changes of ubiquitin binding protein leveljunction protein ubiquitination level. Methods Myocardial cell isolated and subcultured H9c2 were cultured in vitro andmyocardial cell, they were divided into three groups in different Ang Ⅱ concentration（10^（-8）, 10^（-7）, 10^（-6） mol / L）,with in different hours（0, 6, 12, 24, 48, 72 h）, and there is also a control group; measured the total protein contentlevel were measured useby BCA method, measured the number of cardiomyocytes in the same area were measured, and the relative surface area of myocardial cells were calculated. The expression of ubiquitin binding protein cardiomyocyte hypertrophy was time- and concentration-dependently induced by Ang Ⅱ, wheremanifested,as total cellular protein content,relative cell surface area and the expression of ubiquitin binding proteinubiquitinated desmin significantly increasing compared with the control group（P〈0.05）, which were most significant in10^（-7）mol / L AngⅡtreated group for 48 h. Conclusions Cardiomyocyte hypertrophy model induced by 10^（-7） mol / L AngⅡ for 48 h, was successfully established, where H9c2 cardiomyocyte hypertrophy was induced by 10^（-7）mol / L AngⅡ in 48 h, and ubiquitin binding proteinubiquitinated desmin significantly increased in cardiomyocyte hypertrophy.
出处
《实用医学杂志》
CAS
北大核心
2016年第7期1073-1076,共4页
The Journal of Practical Medicine
基金
国家自然科学基金资助项目(编号:81360030)
