Activation of mitogen-activated protein kinases in in vivo ischemia/reperfused myocardium in rats.
In this study, we investigate the in vivo activation of mitogen-activated protein kinases (MAPK) as important signal transduction cascades observed after myocardial ischemia/reperfusion. Myocardial continuous ischemia and ischemia/reperfusion was produced in Wistar rats. The activities of MAPKs in the ischemic and ischemia/reperfused regions were measured using an in-gel kinase assay, an in vitro kinase assay and Western blot analysis. Activator protein-1 (AP-1) DNA binding activity was determined using an electrophoretic mobility shift assay. DNA fragmentation was detected as DNA ladders by agarose gel electrophoresis. The p46JNK and p55JNK activities of continuous ischemia were significantly increased at 30 min (5.9 and 4.2 fold, respectively P<0.05). Coronary reperfusion increased both p42ERK and p44ERK activities at 30 min (3.0 and 2.3 fold P<0.01), and both p46JNK and p55JNK activities at 30 min (1.4 and 1.7 fold P<0.05). The AP-1 DNA binding activities of continuous ischemia were significantly increased at 1, 3 and 7 days (28, 21 and 17 fold, respectively P<0.01). Coronary reperfusion markedly decreased AP-1 DNA binding activities at 1 (41%P<0.01) and 3 days (48%P<0.05). Myocardial DNA fragmentation was considerably more enhanced by reperfusion than continuous ischemia. In conclusion, our present work provides the first in vivo evidence that ERK and JNK are activated by reperfusion from the activities of continuous ischemia. These signal transduction mechanisms may be partially responsible for the myocardial injury.
Omura
T
T
First Department of Internal Medicine, Osaka City University Medical School, Osaka, Japan.
Yoshiyama
M
M
Shimada
T
T
Shimizu
N
N
Kim
S
S
Iwao
H
H
Takeuchi
K
K
Yoshikawa
J
J
D000818:Animals; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism*; D053938:DNA Fragmentation; D004789:Enzyme Activation; D048031:JNK Mitogen-Activated Protein Kinases; D008297:Male; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D017202:Myocardial Ischemia / Q000201:enzymology*; D015425:Myocardial Reperfusion; D010766:Phosphorylation; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction; D018808:Transcription Factor AP-1 / Q000378:metabolism*; D048051:p38 Mitogen-Activated Protein Kinases
D018808:Transcription Factor AP-1; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D048031:JNK Mitogen-Activated Protein Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases