Oxidative stress mediates tumor necrosis factor-α-induced mitochondrial DNA damage and dysfunction in cardiac myocytes

Nobuhiro Suematsu, Hiroyuki Tsutsui, Jing Wen, Dongchon Kang, Masaki Ikeuchi, Tomomi Ide, Shunji Hayashidani, Tetsuya Shiomi, Toru Kubota, Naotaka Hamasaki, Akira Takeshita

Research output: Contribution to journalArticle

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Abstract

Background - Tumor necrosis factor-α (TNF-α) and angiotensin II (Ang II) are implicated in the development and further progression of heart failure, which might be, at least in part, mediated by the production of reactive oxygen species (ROS). However, the cause and consequences of this agonist-mediated ROS production in cardiac myocytes have not been well defined. Recently, we demonstrated that increased ROS production was associated with mitochondrial DNA (mtDNA) damage and dysfunction in failing hearts. We thus investigated whether the direct exposure of cardiac myocytes to TNF-α and Ang II in vitro could induce mtDNA damage via production of ROS. Methods and Results - TNF-α increased ROS production within cultured neonatal rat ventricular myocytes after 1 hour, as assessed by 2′,7′-dichlorofluorescin diacetate fluorescence microscopy. TNF-α also decreased mtDNA copy number by Southern blot analysis in association with complex III activity, which was prevented in the presence of the antioxidant a-tocopherol. A direct exposure of myocytes to H2O2 caused a similar decrease in mtDNA copy number. In contrast, Ang II did not affect mtDNA copy number, despite the similar increase in ROS production. TNF-α-mediated ROS production and a decrease in mtDNA copy number were inhibited by the sphingomyelinase inhibitor D609. Furthermore, N-acetylsphingosine (C2-ceramide), a synthetic cell-permeable ceramide analogue, increased myocyte ROS production, suggesting that TNF-α-mediated ROS production and subsequent mtDNA damage were mediated by the sphingomyelin-ceramide signaling pathway. Conclusions - The intimate link between TNF-α, ROS, and mtDNA damage might play an important role in myocardial remodeling and failure.

Original languageEnglish
Pages (from-to)1418-1423
Number of pages6
JournalCirculation
Volume107
Issue number10
DOIs
Publication statusPublished - Mar 18 2003

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Mitochondrial DNA
Cardiac Myocytes
DNA Damage
Reactive Oxygen Species
Oxidative Stress
Tumor Necrosis Factor-alpha
Angiotensin II
Muscle Cells
Ceramides
Heart Failure
Artificial Cells
Sphingomyelin Phosphodiesterase
Tocopherols
Sphingomyelins
Electron Transport Complex III
Southern Blotting
Fluorescence Microscopy
Antioxidants

All Science Journal Classification (ASJC) codes

  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

Cite this

Oxidative stress mediates tumor necrosis factor-α-induced mitochondrial DNA damage and dysfunction in cardiac myocytes. / Suematsu, Nobuhiro; Tsutsui, Hiroyuki; Wen, Jing; Kang, Dongchon; Ikeuchi, Masaki; Ide, Tomomi; Hayashidani, Shunji; Shiomi, Tetsuya; Kubota, Toru; Hamasaki, Naotaka; Takeshita, Akira.

In: Circulation, Vol. 107, No. 10, 18.03.2003, p. 1418-1423.

Research output: Contribution to journalArticle

Suematsu, Nobuhiro ; Tsutsui, Hiroyuki ; Wen, Jing ; Kang, Dongchon ; Ikeuchi, Masaki ; Ide, Tomomi ; Hayashidani, Shunji ; Shiomi, Tetsuya ; Kubota, Toru ; Hamasaki, Naotaka ; Takeshita, Akira. / Oxidative stress mediates tumor necrosis factor-α-induced mitochondrial DNA damage and dysfunction in cardiac myocytes. In: Circulation. 2003 ; Vol. 107, No. 10. pp. 1418-1423.
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T1 - Oxidative stress mediates tumor necrosis factor-α-induced mitochondrial DNA damage and dysfunction in cardiac myocytes

AU - Suematsu, Nobuhiro

AU - Tsutsui, Hiroyuki

AU - Wen, Jing

AU - Kang, Dongchon

AU - Ikeuchi, Masaki

AU - Ide, Tomomi

AU - Hayashidani, Shunji

AU - Shiomi, Tetsuya

AU - Kubota, Toru

AU - Hamasaki, Naotaka

AU - Takeshita, Akira

PY - 2003/3/18

Y1 - 2003/3/18

N2 - Background - Tumor necrosis factor-α (TNF-α) and angiotensin II (Ang II) are implicated in the development and further progression of heart failure, which might be, at least in part, mediated by the production of reactive oxygen species (ROS). However, the cause and consequences of this agonist-mediated ROS production in cardiac myocytes have not been well defined. Recently, we demonstrated that increased ROS production was associated with mitochondrial DNA (mtDNA) damage and dysfunction in failing hearts. We thus investigated whether the direct exposure of cardiac myocytes to TNF-α and Ang II in vitro could induce mtDNA damage via production of ROS. Methods and Results - TNF-α increased ROS production within cultured neonatal rat ventricular myocytes after 1 hour, as assessed by 2′,7′-dichlorofluorescin diacetate fluorescence microscopy. TNF-α also decreased mtDNA copy number by Southern blot analysis in association with complex III activity, which was prevented in the presence of the antioxidant a-tocopherol. A direct exposure of myocytes to H2O2 caused a similar decrease in mtDNA copy number. In contrast, Ang II did not affect mtDNA copy number, despite the similar increase in ROS production. TNF-α-mediated ROS production and a decrease in mtDNA copy number were inhibited by the sphingomyelinase inhibitor D609. Furthermore, N-acetylsphingosine (C2-ceramide), a synthetic cell-permeable ceramide analogue, increased myocyte ROS production, suggesting that TNF-α-mediated ROS production and subsequent mtDNA damage were mediated by the sphingomyelin-ceramide signaling pathway. Conclusions - The intimate link between TNF-α, ROS, and mtDNA damage might play an important role in myocardial remodeling and failure.

AB - Background - Tumor necrosis factor-α (TNF-α) and angiotensin II (Ang II) are implicated in the development and further progression of heart failure, which might be, at least in part, mediated by the production of reactive oxygen species (ROS). However, the cause and consequences of this agonist-mediated ROS production in cardiac myocytes have not been well defined. Recently, we demonstrated that increased ROS production was associated with mitochondrial DNA (mtDNA) damage and dysfunction in failing hearts. We thus investigated whether the direct exposure of cardiac myocytes to TNF-α and Ang II in vitro could induce mtDNA damage via production of ROS. Methods and Results - TNF-α increased ROS production within cultured neonatal rat ventricular myocytes after 1 hour, as assessed by 2′,7′-dichlorofluorescin diacetate fluorescence microscopy. TNF-α also decreased mtDNA copy number by Southern blot analysis in association with complex III activity, which was prevented in the presence of the antioxidant a-tocopherol. A direct exposure of myocytes to H2O2 caused a similar decrease in mtDNA copy number. In contrast, Ang II did not affect mtDNA copy number, despite the similar increase in ROS production. TNF-α-mediated ROS production and a decrease in mtDNA copy number were inhibited by the sphingomyelinase inhibitor D609. Furthermore, N-acetylsphingosine (C2-ceramide), a synthetic cell-permeable ceramide analogue, increased myocyte ROS production, suggesting that TNF-α-mediated ROS production and subsequent mtDNA damage were mediated by the sphingomyelin-ceramide signaling pathway. Conclusions - The intimate link between TNF-α, ROS, and mtDNA damage might play an important role in myocardial remodeling and failure.

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