CoCl2 induces protective events via the p38-MAPK signalling pathway and ANP in the perfused amphibian heart.
Mitogen-activated protein kinases (MAPKs) constitute one of the most important intracellular signalling pathways. In particular, the p38-MAPK subfamily is known to be activated under various stressful conditions, such as mechanical or oxidative stress. Furthermore, cobalt chloride (CoCl2) has been shown to mimic hypoxic responses in various cell lines and cause overproduction of reactive oxygen species (ROS). In the current study, we investigated the effect of CoCl2 on p38-MAPK signalling pathway in the perfused Rana ridibunda heart. Immunoblot analysis of the phosphorylated, and thus activated, form of p38-MAPK revealed that maximum phosphorylation was attained at 500 micromol l(-1) CoCl2. A similar profile was observed for MAPKAPK2 and Hsp27 phosphorylation (direct and indirect p38-MAPK substrates, respectively). Time course analysis of p38-MAPK phosphorylation pattern showed that the kinase reached its peak within 15 min of treatment with 500 micromol l(-1) CoCl2. Similar results were obtained for Hsp27 phosphorylation. In the presence of the antioxidants Trolox or Lipoic acid, p38-MAPK CoCl2-induced phosphorylation was attenuated. Analogous results were obtained for Hsp27 and MAPKAPK2. In parallel, mRNA levels of the ANP gene, a hormone whose transcriptional regulation has previously been shown to be regulated by p38-MAPK, were examined (semi-quantitative ratiometric RT-PCR). CoCl2 treatment significantly increased ANP mRNA levels, whereas, in the presence of antioxidants, the transcript levels returned to basal values. All the above data indicate that CoCl2 stimulates compensatory mechanisms involving the p38-MAPK signalling cascade along with ANP.
Gaitanaki
Catherine
C
Department of Animal and Human Physiology, School of Biology, University of Athens, Panepistimioupolis, 157 84 Athens, Greece.
Kalpachidou
Theodora
T
Aggeli
Ioanna-Katerina S
IK
Papazafiri
Panagiota
P
Beis
Isidoros
I
D000818:Animals; D009320:Atrial Natriuretic Factor / Q000378:metabolism*; D003035:Cobalt / Q000494:pharmacology*; D006321:Heart / Q000187:drug effects*; D006360:Heat-Shock Proteins / Q000378:metabolism; D066298:In Vitro Techniques; D020935:MAP Kinase Signaling System / Q000187:drug effects*; D009206:Myocardium / Q000378:metabolism*; D010477:Perfusion; D011898:Ranidae / Q000502:physiology*; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
D006360:Heat-Shock Proteins; D003035:Cobalt; D009320:Atrial Natriuretic Factor; D048051:p38 Mitogen-Activated Protein Kinases; C018021:cobaltous chloride