Department of Surgery, Duke University Medical Center, Durham, North Carolina;
Am J Physiol Heart Circ Physiol. 2014 Feb;306(3):H373-81. doi: 10.1152/ajpheart.00411.2013. Epub 2013 Nov 22.
Creatine and phosphocreatine levels are decreased in heart failure, and reductions in myocellular phosphocreatine levels predict the severity of the disease and portend adverse outcomes. Previous studies of transgenic mouse models with increased creatine content higher than two times baseline showed the development of heart failure and shortened lifespan. Given phosphocreatine's role in buffering ATP content, we tested the hypothesis whether elevated cardiac creatine content would alter cardiac function under normal physiological conditions. Here, we report the creation of transgenic mice that overexpress the human creatine transporter (CrT) in cardiac muscle under the control of the α-myosin heavy chain promoter. Cardiac transgene expression was quantified by qRT-PCR, and human CrT protein expression was documented on Western blots and immunohistochemistry using a specific anti-CrT antibody. High-energy phosphate metabolites and cardiac function were measured in transgenic animals and compared with age-matched, wild-type controls. Adult transgenic animals showed increases of 5.7- and 4.7-fold in the content of creatine and free ADP, respectively. Phosphocreatine and ATP levels were two times as high in young transgenic animals but declined to control levels by the time the animals reached 8 wk of age. Transgenic mice appeared to be healthy and had normal life spans. Cardiac morphometry, conscious echocardiography, and pressure-volume loop studies demonstrated mild hypertrophy but normal function. Based on our characterization of the human CrT protein expression, creatine and phosphocreatine content, and cardiac morphometry and function, these transgenic mice provide an in vivo model for examining the therapeutic value of elevated creatine content for cardiac pathologies.
肌酸和磷酸肌酸水平在心力衰竭中降低,肌细胞磷酸肌酸水平的降低预测疾病的严重程度,并预示着不良结局。先前的研究表明,肌酸含量增加超过基线两倍的转基因小鼠模型会发展为心力衰竭并缩短寿命。鉴于磷酸肌酸在缓冲 ATP 含量方面的作用,我们检验了这样一个假设,即在正常生理条件下,升高的心脏肌酸含量是否会改变心脏功能。在这里,我们报告了一种转基因小鼠的创建,该小鼠在α肌球蛋白重链启动子的控制下在心肌中过表达人肌酸转运蛋白(CrT)。通过 qRT-PCR 定量检测心脏转基因表达,并用特异性抗 CrT 抗体在 Western blot 和免疫组织化学上记录人 CrT 蛋白表达。在转基因动物中测量高能磷酸盐代谢物和心脏功能,并与年龄匹配的野生型对照进行比较。成年转基因动物的肌酸和游离 ADP 含量分别增加了 5.7 倍和 4.7 倍。年轻转基因动物的磷酸肌酸和 ATP 水平是对照的两倍,但到动物 8 周龄时降至对照水平。转基因小鼠似乎健康,寿命正常。心脏形态测量、清醒超声心动图和压力-容积环研究表明存在轻度肥大,但功能正常。基于我们对人 CrT 蛋白表达、肌酸和磷酸肌酸含量以及心脏形态和功能的表征,这些转基因小鼠为研究升高的肌酸含量对心脏病变的治疗价值提供了体内模型。