[Effect of calcineurin Aalpha gene overexpression on the myocardium apoptosis induced by hypoxia-reoxygenation and adrenergic receptors].
OBJECTIVE
To investigate the effect of calcineurin AalphacDNA (AdCnAalpha) overexpression as a result of adenovirally mediated gene transfer on neonatal rat cardiac myocyte apoptosis induced by hypoxia-reoxygenation (H/R) and adrenergic receptors.


METHODS
Neonatal rat cardiac myocytes were cultured for 20 h after AdCnAalpha transfection, and treated with isoproterenol (10 micromol/L) and 24 h of hypoxia followed by 4 h of reoxygenation (24H/4R). The cardiac myocyte apoptosis induced by the treatments was assessed by flow cytometry and DNA laddering, and the levels of calcineurin, p38 and phosphorylation p38 (p-p38) were determined by Western blotting and (or) RT-PCR.


RESULTS
AdCnAalpha transfection promoted cultured neonatal rat cardiac myocyte apoptosis induced by isoproterenol+24H/4R as compared with the treated cells without transfection (14.247-/+0.525 vs 10.763-/+1.554, P<0.01), along with greater phosphorylation p38 protein expression (1.60-/+0.22 vs 2.42-/+0.19, P<0.01). The levels of p38 underwent no obvious change after AdCnAalpha transfection (P<0.05).


CONCLUSIONS
AdCnAalpha transfection can promote cardiac myocyte apoptosis induced by H/R and adrenergic receptors, the mechanism of which might be associated with p38 mitongen-activated protein kinase (p38MAPK) activation.
Shen
Xiao-mei
XM
Department of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China. shenxiaomei1030@yahoo.com.cn
Zhang
Ju-yan
JY
Cheng
Bei
B
D000256:Adenoviridae / Q000235:genetics; D000318:Adrenergic beta-Agonists / Q000494:pharmacology; D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D015153:Blotting, Western; D019703:Calcineurin / Q000235:genetics* / Q000378:metabolism; D015687:Cell Hypoxia; D002478:Cells, Cultured; D005260:Female; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics; D007545:Isoproterenol / Q000494:pharmacology; D008297:Male; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010100:Oxygen / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D011941:Receptors, Adrenergic / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
D000318:Adrenergic beta-Agonists; D012333:RNA, Messenger; D011941:Receptors, Adrenergic; D048051:p38 Mitogen-Activated Protein Kinases; D019703:Calcineurin; D007545:Isoproterenol; D010100:Oxygen