Inhibition of hypoxia/reoxygenation-induced apoptosis in metallothionein-overexpressing cardiomyocytes

Guang Wu Wang, Zhanxiang Zhou, Jon B. Klein, Yujian Kang

Research output: Contribution to journalArticle

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Abstract

To study possible mechanisms for metallothionein (MT) inhibition of ischemia-reperfusion-induced myocardial injury, cardiomyocytes isolated from MT-overexpressing transgenic neonatal mouse hearts and nontransgenic controls were subjected to 4 h of hypoxia (5% CO2-95% N2, glucose-free modified Tyrode's solution) followed by 1 h of reoxygenation in MEM + 20% fetal bovine serum (FBS) (5% CO2-95% air), and cytochrome c-mediated caspase-3 activation apoptotic pathway was determined. Hypoxia/reoxygenation-induced apoptosis was significantly suppressed in MT-overexpressing cardiomyocytes, as measured by both terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick-end labeling and annexin V-FITC binding. In association with apoptosis, mitochondrial cytochrome c release, as determined by Western blot, was observed to occur in nontransgenic cardiomyocytes. Correspondingly, caspase-3 was activated as determined by laser confocal microscopic examination with the use of FITC-conjugated antibody against active caspase-3 and by enzymatic assay. The activation of this apoptotic pathway was significantly inhibited in MT-overexpressing cells, as evidenced by both suppression of cytochrome c release and inhibition of caspase-3 activation. The results demonstrate that MT suppresses hypoxia/reoxygenation-induced cardiomyocyte apoptosis through, at least in part, inhibition of cytochrome c-mediated caspase-3 activation.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Heart and Circulatory Physiology
Volume280
Issue number5 49-5
StatePublished - May 1 2001

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Metallothionein
Cardiac Myocytes
Caspase 3
Cytochromes c
Apoptosis
Fluorescein-5-isothiocyanate
Myocardial Reperfusion Injury
DNA Nucleotidylexotransferase
Annexin A5
Enzyme Assays
Transgenic Mice
Lasers
Ischemia
Western Blotting
Air
Hypoxia
Glucose
Antibodies
Serum

All Science Journal Classification (ASJC) codes

  • Physiology
  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

Cite this

Inhibition of hypoxia/reoxygenation-induced apoptosis in metallothionein-overexpressing cardiomyocytes. / Wang, Guang Wu; Zhou, Zhanxiang; Klein, Jon B.; Kang, Yujian.

In: American Journal of Physiology - Heart and Circulatory Physiology, Vol. 280, No. 5 49-5, 01.05.2001.

Research output: Contribution to journalArticle

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