title: Proteome analysis of myocardial tissue following ischemia and reperfusion--effects of complement inhibition.
abstract: Myocardial ischemia-reperfusion injury can be related to complement activation with generation of chemotactic mediators, release of cytokines, leukocyte accumulation, and subsequent severe tissue injury. In this regard, activation of transcription factors (i.e., NFkappaB) and de novo protein synthesis or inflammatory protein degradation seems to play an important role. In the present study, we analyzed the cardiac protein expression following myocardial ischemia (60 min) and reperfusion (180 min) in a rabbit model utilizing two-dimensional electrophoresis and nanoHPLC/ESI-MS/MS for biochemical protein identification. To achieve cardioprotective effects, we used a novel highly selective small molecule C1s inhibitor administered 5 min prior to reperfusion. The reduction of myocardial injury was observed as diminished plasma creatine kinase activity in C1s-INH-248-treated animals (65.2+/-3 vs. 38.5+/-3 U/g protein after 3 h of reperfusion, P<0.05). With proteome analysis we were able to detect 509+/-21 protein spots on the gels of the 3 groups. A pattern of 480 spots with identical positions was found on every gel of myocardial tissue of sham animals, vehicle and C1s-INH-248-treated animals. We analyzed 11 spots, which were identified by mass spectrometry: Superoxide dismutase, alpha-crystallin-chain-B, mitochondrial stress protein, Mn SOD, ATP synthase A chain heart isoform, creatine kinase, and troponin T. All of these proteins were significantly decreased in the vehicle group when we compared to sham-treated animals. Treatment with C1s-INH-248 preserved levels of these proteins. Thus, blocking the classical complement pathway with a highly specific and potent synthetic inhibitor of the activated C1 complex archives cardio-protection by altering and preserving different anti-inflammatory and cytoprotective cascades.
authors.affiliation: Department of Medicine III, Martin-Luther-University, Ernst-Grube-Str. 40, 06097 Halle, Germany. michael.buerke@medizin.uni-halle.de
authors.forename: Michael
authors.initials: M
authors.lastname: Buerke
authors.forename: Hansjörg
authors.initials: H
authors.lastname: Schwertz
authors.forename: Tina
authors.initials: T
authors.lastname: Längin
authors.forename: Ute
authors.initials: U
authors.lastname: Buerke
authors.forename: Roland
authors.initials: R
authors.lastname: Prondzinsky
authors.forename: Herbert
authors.initials: H
authors.lastname: Platsch
authors.forename: Joachim
authors.initials: J
authors.lastname: Richert
authors.forename: Sabine
authors.initials: S
authors.lastname: Bomm
authors.forename: Martin
authors.initials: M
authors.lastname: Schmidt
authors.forename: Heinz
authors.initials: H
authors.lastname: Hillen
authors.forename: Stephan
authors.initials: S
authors.lastname: Lindemann
authors.forename: Gottfried
authors.initials: G
authors.lastname: Blaschke
authors.forename: Ursula
authors.initials: U
authors.lastname: Müller-Werdan
authors.forename: Karl
authors.initials: K
authors.lastname: Werdan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D003174:Complement C1 Inactivator Proteins / Q000494:pharmacology; D003165:Complement System Proteins / Q000187:drug effects / Q000502:physiology*; D003402:Creatine Kinase / Q000032:analysis; D004562:Electrocardiography; D008297:Male; D008869:Microtubule-Associated Proteins / Q000032:analysis; D008969:Molecular Sequence Data; D017202:Myocardial Ischemia / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D015428:Myocardial Reperfusion Injury / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D009206:Myocardium / Q000378:metabolism* / Q000473:pathology; D009336:Necrosis / Q000473:pathology; D009504:Neutrophils / Q000502:physiology; D020543:Proteome / Q000032:analysis*; D011817:Rabbits; D013482:Superoxide Dismutase / Q000032:analysis / Q000737:chemistry; D038203:alpha-Crystallin B Chain / Q000032:analysis
chemical_list: D015415:Biomarkers; C515744:C1s-INH-248; D003174:Complement C1 Inactivator Proteins; D008869:Microtubule-Associated Proteins; D020543:Proteome; D038203:alpha-Crystallin B Chain; D003165:Complement System Proteins; D013482:Superoxide Dismutase; D003402:Creatine Kinase