title: DIGE proteome analysis reveals suitability of ischemic cardiac in vitro model for studying cellular response to acute ischemia and regeneration.
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 by small 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 the cellular 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 were identified 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 human material and will create new possibilities in cardiac research.
authors.affiliation: Division of Molecular Biological-Biochemical Processing Technology, Center for Biotechnology and Biomedicine, Universität Leipzig, Leipzig, Germany.
authors.forename: Sina
authors.initials: S
authors.lastname: Haas
authors.forename: Heinz-Georg
authors.initials: HG
authors.lastname: Jahnke
authors.forename: Nora
authors.initials: N
authors.lastname: Moerbt
authors.forename: Martin
authors.initials: M
authors.lastname: von Bergen
authors.forename: Seyedhossein
authors.initials: S
authors.lastname: Aharinejad
authors.forename: Olena
authors.initials: O
authors.lastname: Andrukhova
authors.forename: Andrea A
authors.initials: AA
authors.lastname: Robitzki
mesh_terms: D000328:Adult; D017209:Apoptosis; D015415:Biomarkers / Q000378:metabolism; D001706:Biopsy; D009202:Cardiomyopathies / Q000473:pathology; D003600:Cytosol / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006321:Heart / Q000502:physiology; D006801:Humans; D008297:Male; D008875:Middle Aged; D017202:Myocardial Ischemia / Q000473:pathology*; D032383:Myocytes, Cardiac / Q000166:cytology; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D012038:Regeneration; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
chemical_list: D015415:Biomarkers; D020543:Proteome