Characterization of the protective effects of cardiotrophin-1 against non-ischemic death stimuli in adult cardiomyocytes.
The aim of this study was to investigate the cytoprotective effects of CT-1 against non-ischemic death stimuli in adult cardiomyocytes. Primary cultures of cardiomyocytes isolated from adult rats were stimulated with either angiotensin II (Ang II) or H(2)O(2) in the presence or absence of CT-1. Cell death was determined by trypan blue exclusion, cell viability by MTT assay and apoptosis by TUNEL-Annexin-V staining. Intracellular pathways were analyzed by the employment of chemical inhibitors and by the assessment of signalling intermediates phosphorylation by Western blot analysis. CT-1 reduced (p<0.01) total cell death and apoptosis induced by either Ang II or H(2)O(2), and increased (p<0.01) cell viability in cardiomyocytes exposed to these stimuli. These effects of CT-1 were abolished in the presence of antibodies specific for gp130 or LIFR and did not require RNA or protein synthesis. Both Wortmannin and PD98059 abolished protective effects of CT-1 against H(2)O(2), whereas only Wortmannin inhibited protection against Ang II. In both cases, Akt kinase activation and Bad phosphorylation were observed. These findings suggest that CT-1 protects adult cardiomyocytes against Ang II- and oxidative stress-induced cell death, via gp130/LIFR and by means of the PI3K/Akt and the p42/44 MAPK intracellular cascades.
López
Natalia
N
Area of Cardiovascular Pathophysiology, School of Medicine, Centre for Applied Medical Research (CIMA), University of Navarra, Avda. Pio XII, 55. 31008, Pamplona, Spain.
Díez
Javier
J
Fortuño
María Antonia
MA
D000375:Aging / Q000502:physiology; D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D002316:Cardiotonic Agents / Q000494:pharmacology*; D002352:Carrier Proteins / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D050822:Cytokine Receptor gp130; D016207:Cytokines / Q000494:pharmacology*; D004268:DNA-Binding Proteins / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D017202:Myocardial Ischemia / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt; D051381:Rats; D017208:Rats, Wistar; D050796:STAT3 Transcription Factor; D015398:Signal Transduction / Q000187:drug effects; D015534:Trans-Activators / Q000378:metabolism; D051025:bcl-Associated Death Protein
D000804:Angiotensin II; D015703:Antigens, CD; D002316:Cardiotonic Agents; D002352:Carrier Proteins; D050822:Cytokine Receptor gp130; D016207:Cytokines; D004268:DNA-Binding Proteins; D006861:Hydrogen Peroxide; D008562:Membrane Glycoproteins; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D019869:Phosphatidylinositol 3-Kinases; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D051057:Proto-Oncogene Proteins c-akt; D050796:STAT3 Transcription Factor; D015534:Trans-Activators; D051025:bcl-Associated Death Protein; C494821:Bad protein, rat; C494201:Il6st protein, rat; C494088:Stat3 protein, rat; C092073:cardiotrophin 1; C494920:Akt1 protein, rat