Inhibition of tumor necrosis factor-α-dependent cardiomyocyte apoptosis by metallothionein

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

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Myocardial cell death is an important cellular event of heart failure. Tumor necrosis factor-α (TNF) accumulates in the failing heart and causes myocyte apoptosis, but the mechanism of this action is unclear. This study was undertaken to examine the relationship between TNF-induced cardiomyocyte apoptosis and activation of p38 mitogen-activated protein kinase (MAPK) through oxidative stress. Primary cultures of neonatal cardiomyocytes isolated from transgenic mouse hearts that overexpress metallothionein (MT) as well as cardiomyocytes isolated from wild-type mice were used. The treatment of wild-type cardiomyocytes with TNF at 10 ng/mL induced apoptosis, as detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling and confirmed by Annexin V-fluorescein isothiocyanate binding. The apoptotic effect of TNF was significantly inhibited in the MT-overexpressing cardiomyocytes. Corresponding to the apoptotic effect, TNF at 10 ng/mL caused rapid phosphorylation of p38 MAPK in wild-type cardiomyocytes. The activation of p38 MAPK was further confirmed by an in vivo experiment treating the mice with TNF and measuring p38 MAPK activity using an immune complex kinase assay. The activation of p38 MAPK was not observed in the MT-overexpressing cardiomyocytes either in vitro or in vivo. Importantly, TNF-induced accumulation of reactive oxygen species was dramatically reduced in the MT-overexpressing cardiomyocytes as determined by a carboxy-H 2 -DCFDA staining method. This study thus suggests that p38 MAPK activation is likely involved in TNF-induced cardiomyocyte apoptosis, which is also related to reactive oxygen species accumulation.

Original languageEnglish (US)
Pages (from-to)209-217
Number of pages9
JournalCardiovascular Toxicology
Volume2
Issue number3
DOIs
StatePublished - Dec 1 2002
Externally publishedYes

Fingerprint

Metallothionein
Cardiac Myocytes
p38 Mitogen-Activated Protein Kinases
Tumor Necrosis Factor-alpha
Apoptosis
Chemical activation
Reactive Oxygen Species
Phosphorylation
Oxidative stress
DNA Nucleotidylexotransferase
Annexin A5
Cell death
Antigen-Antibody Complex
Fluorescein
Labeling
Assays
Muscle Cells
Transgenic Mice
Phosphotransferases
Oxidative Stress

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Toxicology
  • Cardiology and Cardiovascular Medicine

Cite this

Inhibition of tumor necrosis factor-α-dependent cardiomyocyte apoptosis by metallothionein. / Klein, Jon B.; Wang, Guang Wu; Zhou, Zhanxiang; Buridi, Abdul; Kang, Yujian.

In: Cardiovascular Toxicology, Vol. 2, No. 3, 01.12.2002, p. 209-217.

Research output: Contribution to journalArticle

Klein, Jon B. ; Wang, Guang Wu ; Zhou, Zhanxiang ; Buridi, Abdul ; Kang, Yujian. / Inhibition of tumor necrosis factor-α-dependent cardiomyocyte apoptosis by metallothionein. In: Cardiovascular Toxicology. 2002 ; Vol. 2, No. 3. pp. 209-217.
@article{d75c8fd4805b48f4aee2f59cd422f378,
title = "Inhibition of tumor necrosis factor-α-dependent cardiomyocyte apoptosis by metallothionein",
abstract = "Myocardial cell death is an important cellular event of heart failure. Tumor necrosis factor-α (TNF) accumulates in the failing heart and causes myocyte apoptosis, but the mechanism of this action is unclear. This study was undertaken to examine the relationship between TNF-induced cardiomyocyte apoptosis and activation of p38 mitogen-activated protein kinase (MAPK) through oxidative stress. Primary cultures of neonatal cardiomyocytes isolated from transgenic mouse hearts that overexpress metallothionein (MT) as well as cardiomyocytes isolated from wild-type mice were used. The treatment of wild-type cardiomyocytes with TNF at 10 ng/mL induced apoptosis, as detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling and confirmed by Annexin V-fluorescein isothiocyanate binding. The apoptotic effect of TNF was significantly inhibited in the MT-overexpressing cardiomyocytes. Corresponding to the apoptotic effect, TNF at 10 ng/mL caused rapid phosphorylation of p38 MAPK in wild-type cardiomyocytes. The activation of p38 MAPK was further confirmed by an in vivo experiment treating the mice with TNF and measuring p38 MAPK activity using an immune complex kinase assay. The activation of p38 MAPK was not observed in the MT-overexpressing cardiomyocytes either in vitro or in vivo. Importantly, TNF-induced accumulation of reactive oxygen species was dramatically reduced in the MT-overexpressing cardiomyocytes as determined by a carboxy-H 2 -DCFDA staining method. This study thus suggests that p38 MAPK activation is likely involved in TNF-induced cardiomyocyte apoptosis, which is also related to reactive oxygen species accumulation.",
author = "Klein, {Jon B.} and Wang, {Guang Wu} and Zhanxiang Zhou and Abdul Buridi and Yujian Kang",
year = "2002",
month = "12",
day = "1",
doi = "10.1385/CT:2:3:209",
language = "English (US)",
volume = "2",
pages = "209--217",
journal = "Cardiovascular Toxicology",
issn = "1530-7905",
publisher = "Humana Press",
number = "3",

}

TY - JOUR

T1 - Inhibition of tumor necrosis factor-α-dependent cardiomyocyte apoptosis by metallothionein

AU - Klein, Jon B.

AU - Wang, Guang Wu

AU - Zhou, Zhanxiang

AU - Buridi, Abdul

AU - Kang, Yujian

PY - 2002/12/1

Y1 - 2002/12/1

N2 - Myocardial cell death is an important cellular event of heart failure. Tumor necrosis factor-α (TNF) accumulates in the failing heart and causes myocyte apoptosis, but the mechanism of this action is unclear. This study was undertaken to examine the relationship between TNF-induced cardiomyocyte apoptosis and activation of p38 mitogen-activated protein kinase (MAPK) through oxidative stress. Primary cultures of neonatal cardiomyocytes isolated from transgenic mouse hearts that overexpress metallothionein (MT) as well as cardiomyocytes isolated from wild-type mice were used. The treatment of wild-type cardiomyocytes with TNF at 10 ng/mL induced apoptosis, as detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling and confirmed by Annexin V-fluorescein isothiocyanate binding. The apoptotic effect of TNF was significantly inhibited in the MT-overexpressing cardiomyocytes. Corresponding to the apoptotic effect, TNF at 10 ng/mL caused rapid phosphorylation of p38 MAPK in wild-type cardiomyocytes. The activation of p38 MAPK was further confirmed by an in vivo experiment treating the mice with TNF and measuring p38 MAPK activity using an immune complex kinase assay. The activation of p38 MAPK was not observed in the MT-overexpressing cardiomyocytes either in vitro or in vivo. Importantly, TNF-induced accumulation of reactive oxygen species was dramatically reduced in the MT-overexpressing cardiomyocytes as determined by a carboxy-H 2 -DCFDA staining method. This study thus suggests that p38 MAPK activation is likely involved in TNF-induced cardiomyocyte apoptosis, which is also related to reactive oxygen species accumulation.

AB - Myocardial cell death is an important cellular event of heart failure. Tumor necrosis factor-α (TNF) accumulates in the failing heart and causes myocyte apoptosis, but the mechanism of this action is unclear. This study was undertaken to examine the relationship between TNF-induced cardiomyocyte apoptosis and activation of p38 mitogen-activated protein kinase (MAPK) through oxidative stress. Primary cultures of neonatal cardiomyocytes isolated from transgenic mouse hearts that overexpress metallothionein (MT) as well as cardiomyocytes isolated from wild-type mice were used. The treatment of wild-type cardiomyocytes with TNF at 10 ng/mL induced apoptosis, as detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling and confirmed by Annexin V-fluorescein isothiocyanate binding. The apoptotic effect of TNF was significantly inhibited in the MT-overexpressing cardiomyocytes. Corresponding to the apoptotic effect, TNF at 10 ng/mL caused rapid phosphorylation of p38 MAPK in wild-type cardiomyocytes. The activation of p38 MAPK was further confirmed by an in vivo experiment treating the mice with TNF and measuring p38 MAPK activity using an immune complex kinase assay. The activation of p38 MAPK was not observed in the MT-overexpressing cardiomyocytes either in vitro or in vivo. Importantly, TNF-induced accumulation of reactive oxygen species was dramatically reduced in the MT-overexpressing cardiomyocytes as determined by a carboxy-H 2 -DCFDA staining method. This study thus suggests that p38 MAPK activation is likely involved in TNF-induced cardiomyocyte apoptosis, which is also related to reactive oxygen species accumulation.

UR - http://www.scopus.com/inward/record.url?scp=0038506234&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0038506234&partnerID=8YFLogxK

U2 - 10.1385/CT:2:3:209

DO - 10.1385/CT:2:3:209

M3 - Article

C2 - 12665666

AN - SCOPUS:0038506234

VL - 2

SP - 209

EP - 217

JO - Cardiovascular Toxicology

JF - Cardiovascular Toxicology

SN - 1530-7905

IS - 3

ER -