血管紧张素Ⅱ联合缺氧诱导大鼠心肌细胞肥大的模型制备及应用  

Model preparation and Application of cardiomyocyte hypertrophy induced by Ang Ⅱ and anoxia

作　　者：李芳 杜小换[1] 王文静 朱增燕[1] Li Fang;Du Xiaohuan;Wang Wenjing;Zhu Zengyan(Department of Pharmacy,Children's Hospital affiliated to Soochow University,Suzhou,215025,China;不详)

机构地区：[1]苏州大学附属儿童医院药剂科,苏州215025

出　　处：《中国循证心血管医学杂志》2020年第12期1475-1479,1482,共6页Chinese Journal of Evidence-Based Cardiovascular Medicine

基　　金：江苏省苏州市民生科技项目(SYS2018073)。

摘　　要：目的采用血管紧张素Ⅱ(AngⅡ)联合缺氧制备大鼠心肌细胞肥大模型并探讨其应用。方法采用不同浓度AngⅡ作用不同时间和/或不同时间缺氧处理诱导大鼠心肌细胞H9c2(2-1)肥大,观察心肌细胞存活率,测定培养上清中乳酸脱氢酶(LDH)活性、细胞内总蛋白和肥大标记物心钠肽(ANP)含量,初步确定AngⅡ作用浓度和时间;进一步检测细胞内缺氧诱导因子-1α(HIF-1α)蛋白表达,再结合培养上清中LDH的活性,最终确定最佳造模条件。通过HIF-1α抑制剂牡荆素作用肥大H9c2细胞后检测胞内ANP和游离脂肪酸(FFA)含量、测定细胞内HIF-1α蛋白表达,培养上清中葡萄糖水平来进行模型应用。结果10-6 mol/L AngⅡ作用24 h联合缺氧4 h可诱导大鼠心肌细胞肥大。牡荆素可降低肥大H9c2细胞内ANP、FFA含量及HIF-1α蛋白表达,提高培养液上清中葡萄糖水平。结论AngⅡ联合缺氧培养可成功制备大鼠心肌细胞肥大模型,该模型可应用于抗心肌肥厚药物的体外研究。Objective AngiotensinⅡ(ANGⅡ)combined with hypoxia was used to prepare a rat cardiomyocyte hypertrophy model and explore its application.Method H9c2(2-1)hypertrophy was induced in rat cardiac myocytes by treatment with different concentrations of AngⅡfor different time and/or hypoxia treatment for different time,cell viability was observed,lactate dehydrogenase(LDH)activity in the culture supernatant,total intracellular protein and the hypertrophy marker atrial natriuretic peptide(ANP)content were measured,and the concentration and time of AngⅡaction were initially determined;the induction of hypoxia in the cells was further examinedSub-1α(HIF-1α)protein expression,combined with LDH activity in the culture supernatant,to ultimately determine the optimal modeling conditions.The model application was performed by measuring the intracellular ANP and free fatty acid(FFA)contents,measuring the intracellular HIF-1αprotein expression,and determining the glucose level in the culture supernatant after the hypertrophic H9c2 cells were treated with Vitexin,a HIF-1αinhibitor.Result 10-6 mol/L AngⅡfor 24 h combined with hypoxia for 4 h induced hypertrophy of rat cardiomyocytes.Vitexin decreased ANP and FFA contents and HIF-1αprotein expression in hypertrophic H9c2 cells and increased glucose levels in the supernatant.Conclusion AngⅡcombined with hypoxia culture can successfully prepare rat cardiomyocyte hypertrophy model,and this model can be applied to the study of anti cardiac hypertrophic drugs in vitro.

关 键 词：血管紧张素Ⅱ 缺氧培养 心肌细胞肥大 模型制备 应用 

分 类 号：R542.2[医药卫生—心血管疾病]