Isquemia y regeneracion

DIGE Proteome Analysis Reveals Suitability of Ischemic Cardiac In Vitro Model for Studying Cellular Response to Acute Ischemia and Regeneration
Sina Haas1, Heinz-Georg Jahnke1, Nora Moerbt2, Martin von Bergen2,3, Seyedhossein Aharinejad4,5, Olena Andrukhova5,6, Andrea A. Robitzki1*
¨ 1 Division of Molecular Biological-Biochemical Processing Technology, Center for Biotechnology and Biomedicine,Universitat Leipzig, Leipzig, Germany, 2 Department of Proteomics, Helmholtz Centre for Environmental Research, Leipzig, Germany, 3 Department of Metabolomics, Helmholtz Centre for Environmental Research, Leipzig, Germany, 4 Department of Cardiac Surgery, Center of Anatomy and Cell Biology, Medical University of Vienna, Vienna, Austria, 5 Department for Cardiovascular Research, Center of Anatomyand Cell Biology, Medical University of Vienna, Vienna, Austria, 6 Department for Biomedical Sciences, Institute of Pathophysiology, University of Veterinary Medicine, Vienna, Austria

Abstract
Proteomic analysis of myocardial tissue from patient population is suited to yield insights into cellular and molecular mechanisms taking place in cardiovascular diseases. However, it has been limited bysmall sized biopsies and complicated by high variances between patients. Therefore, there is a high demand for suitable model systems with the capability to simulate ischemic and cardiotoxic effects in vitro, under defined conditions. In this context, we established an in vitro ischemia/reperfusion cardiac disease model based on the contractile HL-1 cell line. To identify pathways involved in thecellular alterations induced by ischemia and thereby defining disease-specific biomarkers and potential target structures for new drug candidates we used fluorescence 2D-difference gel electrophoresis. By comparing spot density changes in ischemic and reperfusion samples we detected several protein spots that were differentially abundant. Using MALDI-TOF/ TOF-MS and ESI-MS the proteins wereidentified and subsequently grouped by functionality. Most prominent were changes in apoptosis signalling, cell structure and energy-metabolism. Alterations were confirmed by analysis of human biopsies from patients with ischemic cardiomyopathy. With the establishment of our in vitro disease model for ischemia injury target identification via proteomic research becomes independent from rare humanmaterial and will create new possibilities in cardiac research.
Citation: Haas S, Jahnke H-G, Moerbt N, von Bergen M, Aharinejad S, et al. (2012) DIGE Proteome Analysis Reveals Suitability of Ischemic Cardiac In Vitro Model for Studying Cellular Response to Acute Ischemia and Regeneration. PLoS ONE 7(2): e31669. doi:10.1371/journal.pone.0031669 ˜o Editor: Rodrigo Alexandre Panepucci, Hemocentro deRibeira Preto - HC-FMRP-USP., Brazil Received July 18, 2011; Accepted January 11, 2012; Published February 22, 2012 Copyright: ß 2012 Haas et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work wasfunded by the European Union (FP6, Project: CWB 266753) and the BuildMoNa Graduate School of Leipzig. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. * E-mail: andrea.robitzki@bbz.uni-leipzig.de

Introduction
Cardiac diseases andmyocardial dysfunctions following ischemia are the leading cause of mortality in western industrialized countries. Ischemia and reperfusion injury, resulting from clinical setting of coronary revascularization in acute myocardial infarction, bypass surgery and heart transplantation is a demanding issue. Many dysfunctions and defects have been described to be responsible for the occurrence of ischemic...