Division of Thoracic & Cardiovascular Surgery, Chang Gung Memorial Hospital at Keelung, Taiwan.
Shock. 2012 Dec;38(6):607-14. doi: 10.1097/SHK.0b013e318271f944.
In this study, experiments were designed to determine whether microRNAs (miRNAs) play a role in the regulation or modulation of cardiomyocytic reactions under cardioplegia-induced cardiac arrest during cardiopulmonary bypass. MicroRNAs play powerful and unexpected roles in numerous cardiovascular diseases. MicroRNA-based therapeutics may provide a unique opportunity to translate this knowledge into the clinical setting. Sprague-Dawley rats (10 per group) were randomly divided into three groups: control, perfusion, and arrest groups. In the perfusion group, isolated hearts were perfused with oxygenated physiologic buffered solution for 3 h using a Langendorff apparatus. In the arrest group, cold crystalloid cardioplegia solution was used to induce and maintain cardiac arrest for 1 h; hearts were reperfused for 2 h with warm oxygenated phosphate-buffered saline solution. Cardiac miRNAs and protein expression patterns were detected using miRNA arrays and two-dimensional fluorescence difference gel electrophoresis followed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Of 103 different miRNAs and 222 different proteins expressed in the three groups, miRNA-27a was the one considered to be related to the regulation of cardiomyocyte apoptosis by targeting the interleukin 10 pathway. Transfection of H9c2 cardiomyocytes with pre-miRNA-27a, which significantly decreased the mRNA and protein levels of interleukin 10 and increased expression of nuclear factor κB and its downstream cytokines during hypoxia/reperfusion injury, could activate caspase 3 and apoptosis. Our study demonstrated the altered expression of miRNAs in cardiomyocytes during cardioplegia-induced cardiac arrest. The involvement of miRNAs in cardiomyocytic apoptosis adds another level of complexity to gene regulation, which could open up novel avenues for cardiac protection strategies during cardiac surgery.
在这项研究中,设计了实验来确定 microRNAs(miRNAs)是否在心肺旁路期间心脏停搏时心脏停搏诱导的心肌反应的调节或调制中发挥作用。miRNAs 在许多心血管疾病中发挥强大且出乎意料的作用。基于 miRNA 的治疗方法可能为将这一知识转化为临床环境提供独特的机会。10 只大鼠(每组 10 只)被随机分为三组:对照组、灌注组和停搏组。在灌注组中,使用 Langendorff 仪器用含氧生理缓冲液对分离的心脏进行灌注 3 小时。在停搏组中,使用冷结晶心脏停搏液诱导和维持心脏停搏 1 小时;用温热的含氧磷酸盐缓冲盐水溶液再灌注 2 小时。使用 miRNA 阵列和二维荧光差异凝胶电泳后,通过基质辅助激光解吸/电离飞行时间质谱法检测心脏中的 miRNA 和蛋白质表达模式。在三组中表达的 103 种不同的 miRNA 和 222 种不同的蛋白质中,miRNA-27a 被认为与通过靶向白细胞介素 10 途径调节心肌细胞凋亡有关。转染 H9c2 心肌细胞的 pre-miRNA-27a 可显著降低白细胞介素 10 的 mRNA 和蛋白质水平,并在缺氧/再灌注损伤期间增加核因子 κB 及其下游细胞因子的表达,可激活 caspase 3 和细胞凋亡。我们的研究表明,在心脏停搏诱导的心脏停搏期间,心肌细胞中 miRNA 的表达发生改变。miRNAs 参与心肌细胞凋亡为基因调控增加了另一个层次的复杂性,这可能为心脏手术期间的心脏保护策略开辟新途径。