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Establishment of an easy and straight forward heparinase protocol to analyse circulating and myocardial tissue micro-RNA during coronary artery-bypass-graft surgery

机译:建立简单直接的肝素酶方案以分析冠状动脉搭桥手术中的循环和心肌组织微小RNA

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摘要

Coronary artery-bypass-graft (CABG) surgery is associated with myocardial damage and increased blood concentrations of circulating microRNAs (miRNA). However, whether and to what extent these miRNAs relate to cardiac tissue miRNA expression have not yet been explored. Since plasma miRNA quantification in samples from cardiopulmonary bypass (CPB) patients is severely hampered by heparin, we established and validated successfully a protocol to reliably measure miRNA in 49 heparinized patients undergoing CABG so as to investigate the relationship between circulating and right atrial miRNAs. Plasma and right atrial expression of miR-1, miR-133a, miR-423-5p, and miR-499 were measured before and after CPB, as well as miRNAs in plasma 24 h thereafter. All plasma miRNAs increased significantly with surgery while cardiac tissue expression of only miR-133a (1.4-fold; p = 0.003) and miR-423-5p (1.3 fold; p = 0.025) increased as well. Right atrial and plasma miR-133a expression correlated positively before CPB (r = 0.288, p = 0.045) but miR-499 expression inversely (r = −0.484, p = 0.0004). There was a strong association between plasma miR-133a and miR-499 concentrations and postoperative troponin I concentrations, the marker for myocardial damage. Increased myocardial miR-133a and miR-423-5p expression together with unchanged miR-1 and miR-499 expression might suggest active release of these miRNAs rather than their origin from damaged cells.
机译:冠状动脉旁路移植术(CABG)与心肌损伤和循环微RNA(miRNA)血液浓度升高有关。然而,尚未探究这些miRNA是否与心脏组织miRNA表达有关以及在何种程度上与心脏组织miRNA表达有关。由于肝素严重阻碍了体外循环(CPB)患者样本中血浆miRNA的定量分析,因此我们建立并成功验证了一项可靠的方案,可以可靠地测量49位接受CABG的肝素化患者中的miRNA,从而研究循环性和右房性miRNA之间的关系。在CPB前后分别测量miR-1,miR-133a,miR-423-5p和miR-499的血浆和右心房表达,以及之后24h血浆中的miRNA。所有血浆miRNA随手术而显着增加,而心脏组织中仅miR-133a(1.4倍; p = 0.003)和miR-423-5p(1.3倍; p = 0.025)的表达也增加。右心房和血浆miR-133a表达在CPB之前呈正相关(r = 0.288,p = 0.045),而miR-499表达相反(r = -0.484,p = 0.0004)。血浆miR-133a和miR-499的浓度与术后肌钙蛋白I的浓度(心肌损伤的标志物)之间存在很强的联系。心肌miR-133a和miR-423-5p的表达增加,以及miR-1和miR-499的表达不变可能表明这些miRNA的主动释放,而不是它们源自受损细胞的起源。

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