Blood mRNA Expression Profiles of Autophagy, Apoptosis, and Hypoxia Markers on Blood Cardioplegia and Custodiol Cardioplegia Groups

Abstract Introduction: Blood cardioplegia (BC) and Custodiol cardioplegia (CC) have been used for a long time in open heart surgery and are highly effective solutions. The most controversial issue among these two is whether there is any difference between them regarding myocardial damage after ischemia surgery. In this study, autophagy, apoptosis, and hypoxia markers were investigated and that way we evaluated the differences between BC and CC patients. Methods: A total of 30 patients were included in this study, using two different cardioplegic solutions. Three different whole blood samples of the patients were taken from a central vein (preoperatively, immediately postoperatively, and one day after surgery). Total ribonucleic acid was extracted from these samples. Quantitative real-time polymerase chain reaction was performed, and changes in gene expression were determined by the 2-∆∆Ct method of relative quantification. Results: In the CC group, Beclin gene expression level was found to be higher and this difference was statistically significant (P=0.0024). Similarly, cysteine-aspartic acid protease (caspase) 9 and hypoxia-inducible factor 1α messenger ribonucleic acid (mRNA) gene expression level increased and were significantly different in the CC group. In the BC group, Beclin and microtubule-associated protein light chain 3 expressions were higher in the samples taken one day after surgery. Caspases 3 and 8 gene expressions were significantly different in the BC group. Conclusion: As a result of the analysis performed between the two cardioplegia groups, it has been shown that CC harms the myocardium more than BC at the level of mRNA expression of related markers.

摘要 引言：血液心脏停搏液（Blood cardioplegia, BC）与库司迪尔心脏停搏液（Custodiol cardioplegia, CC）均已在心脏直视手术中应用多年，且为高效的心脏停搏液制剂。二者之间最具争议的问题在于，缺血术后心肌损伤方面是否存在差异。本研究通过检测自噬（autophagy）、凋亡（apoptosis）及缺氧（hypoxia）相关标志物，对接受两种心脏停搏液治疗的患者差异进行评估。
方法：本研究共纳入30例患者，分别使用两种不同的心脏停搏液。分别于术前、术后即刻及术后1天，从患者中心静脉采集全血样本，共3份。从上述样本中提取总核糖核酸（ribonucleic acid, RNA），采用实时荧光定量聚合酶链反应（quantitative real-time polymerase chain reaction, qRT-PCR）进行检测，并通过2^(-ΔΔCt)相对定量法分析基因表达变化。
结果：CC组患者的Beclin1基因表达水平更高，差异具有统计学意义（P=0.0024）。同理，CC组患者的半胱天冬氨酸蛋白酶（cysteine-aspartic acid protease, caspase）9及缺氧诱导因子1α信使核糖核酸（messenger ribonucleic acid, mRNA）的基因表达水平均显著升高。BC组患者术后1天采集的样本中，Beclin1及微管相关蛋白轻链3（microtubule-associated protein light chain 3, LC3）的表达水平更高。BC组患者的胱天蛋白酶3、8基因表达差异同样具有统计学意义。
结论：对两组心脏停搏液患者的分析结果显示，在相关标志物的mRNA表达层面，CC对心肌的损伤程度高于BC。