摘要
目的研究转化生长因子β1(TGF-β1)对血管紧张素Ⅱ(AngⅡ)作用后的心肌细胞表面积和心钠素合成影响。方法分离培养原代心肌细胞,分为5组,一组常规培养,一组采用AngⅡ处理,一组采用TGF-β1抑制剂和AngⅡ共同处理心肌细胞(并根据TGF-β1不同浓度分为3组),实时荧光定量聚合酶链反应(RT-PCR)和免疫印迹法(Western blot)分别检测细胞中TGF-β1 mRNA和蛋白表达。测定心肌细胞表面积,免疫放射法测定心钠素合成情况,硝酸还原法测定一氧化氮(NO)合成情况,试剂盒测定细胞中的蛋白含量,流式细胞术测定细胞凋亡情况,Western blot法测定各组细胞中B细胞淋巴瘤/白血病-2(Bcl-2)、细胞内信号转导蛋白2(smad2)表达。结果 AngⅡ处理后的心肌细胞中TGF-β1 mRNA和蛋白水平均明显高于常规培养的心肌细胞(P<0.05)。AngⅡ作用后的心肌细胞凋亡率升高,Bcl-2蛋白水平下降,smad2蛋白水平升高,细胞表面积明显增加,并且心肌细胞合成的心钠素水平也明显升高,心肌细胞蛋白含量也明显升高,而心肌细胞合成的NO含量下降,与常规培养的心肌细胞相比差异具有统计学意义(P<0.05)。用不同浓度的TGF-β1抑制剂作用后的心肌细胞凋亡率降低,Bcl-2蛋白水平升高,smad2蛋白水平下降,细胞表面积有所降低,细胞合成的心钠素减少,心肌细胞蛋白含量也下降,心肌细胞合成的NO水平升高,与AngⅡ作用后的心肌细胞相比差异具有统计学意义(P<0.05)。结论 TGF-β1在AngⅡ作用后的心肌细胞中高表达,抑制TGF-β1可以减弱AngⅡ诱导的心肌细胞肥大,这可能与抑制心肌细胞凋亡和促进NO合成有关。
Objective To study the influence of transforming growth factorβ1(TGF-β1)on surface area of myocardiocytes and synthesis of atrial natriuretic peptide(ANP)after effected by angiotensin II(Ang II).Methods Primary myocardiocytes were isolated,cultured and divided into 5 groups:control group(normally cultured),Ang II group(treated with Ang II),and test groups 1,2 and 3(treated with Ang II and TGF-β1 in different doses).The mRNA and protein expressions of TGF-β1 were detected by using real-time fluorescence quantitative polymerase chain reaction(RT-PCR).The surface area of myocardiocytes was determined,and synthesis of ANP was detected by using radioimmunoassay.The synthesis of nitric oxide(NO)was detected by using nitrate reductase method,and protein content in myocardiocytes was determined by using kit.The apoptosis of myocardiocytes was detected by using flow cytometry.The expressions of Bcl-2 and smad2 were detected by using Western blotting assay in all groups.Results The mRNA and protein expressions of TGF-β1 were significantly higher in Ang II group than those in control group(P<0.05).The apoptosis rate of myocardiocytes increased,Bcl-2 level decreased,smad2 level increased,surface area of myocardiocytes increased significantly,ANP level increased significantly,protein content in myocardiocytes increased significantly,and NO level decreased in Ang II group compared with control group(P<0.05).The apoptosis rate of myocardiocytes decreased,Bcl-2 level increased,smad2 level decreased,surface area of myocardiocytes decreased,ANP level decreased,protein content in myocardiocytes decreased,and NO level increased in test groups 1,2 and 3 compared with Ang II group(P<0.05).Conclusion TGF-β1 has a higher expression in myocardiocytes affected with Ang II.Inhibition of TGF-β1 can relieve cardiomyocyte hypertrophy induced by Ang II,which may be related to inhibition of cardiomyocyte apoptosis and NO synthesis.
作者
聂明
金培印
Nie Ming;Jin Peiyin(Department of Cardiology,People's Hospital of Anyang City,Anyang 455000,China)
出处
《中国循证心血管医学杂志》
2018年第10期1254-1257,1260,共5页
Chinese Journal of Evidence-Based Cardiovascular Medicine
关键词
转化生长因子Β1
心肌细胞肥大
血管紧张素Ⅱ
心钠素
Transforming growth factorβ1
Cardiomyocyte hypertrophy
AngiotensinⅡ
Atrial natriuretic peptide
