摘要
目的检测TNNI3K基因过表达对成年大鼠心肌细胞肌丝钙敏感度变化的影响,为进一步研究TNNI3K基因在心肌收缩功能、心肌肥厚、心衰及其它心脏疾病中的作用提供了可靠有效的研究模型。方法恒压Langedoff灌流装置逆向灌流法分离成年大鼠心肌细胞,之后对该心肌细胞进行逐级复钙。然后,将心肌细胞接种于改良的M199培养基中。分别感染Ad.GFP病毒和Ad.TNNI3K病毒,24小时后,以IonOptix单细胞可视化动缘系统检测心肌细胞肌丝钙敏感度。结果成功分离培养了成年大鼠心肌细胞,并在该细胞中成功过表达了TNNI3K基因。钙敏感度测定结果表明,TNNI3K基因能够增强心肌细胞肌丝钙敏感度。结论在本实验成功分离培养的成年大鼠心肌细胞中以腺病毒载体过表达TNNI3K基因,可增强心肌细胞肌丝的钙敏感度,该模型为进一步研究TNNI3K基因在心肌收缩功能中的作用及机制奠定了基础。
Objective To provide a useful model for studying the function of TNNI3K during the heart contraction, hypertrophy, heart failure and other heart diseases, adult rat cardiac myocytes were cultured and the effect of TNNI3K overexpression on Ca2+ sensitivity was determined. Methods Sprague-Dawley male rats were anesthetized and the heart was quickly removed from the chest. Then the heart was retroperfused with an enzyme solution containing collegenase II. After digestion was terminated, left ventricular tissue...
出处
《中国分子心脏病学杂志》
CAS
2011年第2期83-87,共5页
Molecular Cardiology of China
基金
国家自然科学基金自助项目(30871057)
