Witort Ewa, Capaccioli Sergio, Becatti Matteo, Fiorillo Claudia, Batignani Giacomo, Pavoni Vittorio, Piccini Matteo, Orioli Marica, Carini Marina, Aldini Giancarlo, Lulli Matteo
a Department of Experimental and Clinical Biomedical Sciences , University of Florence , Florence , Italy ;
b Department of Surgery and Translational Medicine , University of Florence , Florence , Italy ;
Free Radic Res. 2016 Aug;50(8):831-9. doi: 10.1080/10715762.2016.1179736. Epub 2016 May 12.
The aim of this study was to measure and identify the reactive carbonyl species (RCSs) released in the blood of humans subjected to hepatic resection. Pre-anesthesia malondialdehyde (MDA) plasma content (0.36 ± 0.11 nmol/mg protein) remained almost unchanged immediately after anaesthesia, before clamping and at the 10th min after ischemia, while markedly increased (to 0.59 ± 0.07 nmol/mg; p < 0.01, Tukey's post test) at the 10th min of reperfusion. A similar trend was observed for the protein carbonyls (PCs), whose pre-anesthesia levels (0.17 ± 0.13 nmol/mg) did not significantly change during ischemia, while increased more than fourfold at the 10th min of reperfusion (0.75 ± 0.17 nmol/mg; p < 0.01, Tukey's post test). RCSs were then identified as covalent adducts to the albumin Cys34, which we previously found as the most reactive protein nucleophilic site in plasma. By using a mass spectrometry (MS) approach based on precursor ion scanning, we found that acrolein (ACR) is the main RCS adducted to albumin Cys34. In basal conditions, the adducted albumin was 0.6 ± 0.4% of the native form but it increased by almost fourfold at the 10th min of reperfusion (2.3 ± 0.7%; p < 0.01, t-test analysis). Since RCSs are damaging molecules, we propose that RCSs, and ACR in particular, are new targets for novel molecular treatments aimed at reducing the ischemia/reperfusion damage by the use of RCS sequestering agents.
本研究的目的是测量和鉴定肝切除患者血液中释放的反应性羰基化合物（RCSs）。麻醉前丙二醛（MDA）血浆含量（0.36±0.11 nmol/mg蛋白质）在麻醉后、夹闭前和缺血后第10分钟时几乎保持不变，而在再灌注第10分钟时显著增加（至0.59±0.07 nmol/mg；p<0.01，Tukey事后检验）。蛋白质羰基（PCs）也观察到类似趋势，其麻醉前水平（0.17±0.13 nmol/mg）在缺血期间无显著变化，而在再灌注第10分钟时增加了四倍多（0.75±0.17 nmol/mg；p<0.01，Tukey事后检验）。然后将RCSs鉴定为与白蛋白Cys34的共价加合物，我们之前发现它是血浆中最具反应性的蛋白质亲核位点。通过基于前体离子扫描的质谱（MS）方法，我们发现丙烯醛（ACR）是与白蛋白Cys34加合的主要RCS。在基础条件下，加合白蛋白占天然形式的0.6±0.4%，但在再灌注第10分钟时增加了近四倍（2.3±0.7%；p<0.01，t检验分析）。由于RCSs是具有破坏性的分子，我们提出RCSs，尤其是ACR，是新型分子治疗的新靶点，旨在通过使用RCS螯合剂减少缺血/再灌注损伤。
