Abstract

Background: Clinical use of doxorubicin is greatly limited by its severe cardiotoxic side effects. L-carnitine is a vitamin-like substance which has been successfully used in many cardiomyopathies, however, the intracellular mechanism(s) remain unclear. The objective of this study was set to evaluate the protective effect of L-carnitine on doxorubicin-induced cardiomyocyte apoptosis, and to explore its intracellular mechanism(s). Methods: Primary cultured neonatal rat cardiomyocytes were treated with doxorubicin (1 μM) with or without pretreatment with L-carnitine (1-30 mM). Lactate dehydrogenase assay, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling staining, and flow cytometry measurement were used to assess cytotoxicity and apoptosis. Fluorescent probes 2',7'-dichlorofluorescein diacetate and chemiluminescence assay of superoxide production were used to detect the production of reactive oxygen species. Western blotting was used to evaluate the quantity of cleaved caspase-3, cytosol cytochrome c, and Bcl-x L expression. Results: L-carnitine inhibited doxorubicin-induced reactive oxygen species generation and NADPH oxidase activation, reduced the quantity of cleaved caspase-3 and cytosol cytochrome c, and increased Bcl-x L expression, resulting in protecting cardiomyocytes from doxorubicin-induced apoptosis. In addition, L-carnitine was found to increase the prostacyclin (PGI2) generation in cardiomyocytes. The siRNA transfection for PGI2 synthase significantly reduced L-carnitine-induced PGI2 and L-carnitine's protective effect. Furthermore, blockade the potential PGI2 receptors, including PGI2 receptors (IP receptors), and peroxisome proliferator-activated receptors alpha and delta (PPARα and PPARδ), revealed that the siRNA-mediated blockage of PPARα considerably reduced the anti-apoptotic effect of L-carnitine. Conclusions: These findings suggest that L-carnitine protects cardiomyocytes from doxorubicin-induced apoptosis in part through PGI2 and PPARα-signaling pathways, which may potentially protect the heart from the severe toxicity of doxorubicin.

Original languageEnglish
Pages (from-to)145-152
Number of pages8
JournalInternational Journal of Cardiology
Volume146
Issue number2
DOIs
Publication statusPublished - Jan 21 2011

Fingerprint

Carnitine
Epoprostenol
Cardiac Myocytes
Doxorubicin
Apoptosis
Peroxisome Proliferator-Activated Receptors
Epoprostenol Receptors
Cytochromes c
Caspase 3
Cytosol
Small Interfering RNA
Reactive Oxygen Species
PPAR delta
PPAR alpha
DNA Nucleotidylexotransferase
NADPH Oxidase
Luminescence
Cardiomyopathies
Fluorescent Dyes
L-Lactate Dehydrogenase

Keywords

  • Apoptosis
  • Cardiomyocytes
  • Doxorubicin
  • L-carnitine
  • Peroxisome proliferator-activated receptor alpha (PPAR)
  • Prostacyclin (PGI )

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

L-carnitine reduces doxorubicin-induced apoptosis through a prostacyclin-mediated pathway in neonatal rat cardiomyocytes. / Chao, Hung Hsin; Liu, Ju Chi; Hong, Hong Jye; Lin, Jia Wei; Chen, Cheng Hsien; Cheng, Tzu-Hurng.

In: International Journal of Cardiology, Vol. 146, No. 2, 21.01.2011, p. 145-152.

Research output: Contribution to journalArticle

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abstract = "Background: Clinical use of doxorubicin is greatly limited by its severe cardiotoxic side effects. L-carnitine is a vitamin-like substance which has been successfully used in many cardiomyopathies, however, the intracellular mechanism(s) remain unclear. The objective of this study was set to evaluate the protective effect of L-carnitine on doxorubicin-induced cardiomyocyte apoptosis, and to explore its intracellular mechanism(s). Methods: Primary cultured neonatal rat cardiomyocytes were treated with doxorubicin (1 μM) with or without pretreatment with L-carnitine (1-30 mM). Lactate dehydrogenase assay, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling staining, and flow cytometry measurement were used to assess cytotoxicity and apoptosis. Fluorescent probes 2',7'-dichlorofluorescein diacetate and chemiluminescence assay of superoxide production were used to detect the production of reactive oxygen species. Western blotting was used to evaluate the quantity of cleaved caspase-3, cytosol cytochrome c, and Bcl-x L expression. Results: L-carnitine inhibited doxorubicin-induced reactive oxygen species generation and NADPH oxidase activation, reduced the quantity of cleaved caspase-3 and cytosol cytochrome c, and increased Bcl-x L expression, resulting in protecting cardiomyocytes from doxorubicin-induced apoptosis. In addition, L-carnitine was found to increase the prostacyclin (PGI2) generation in cardiomyocytes. The siRNA transfection for PGI2 synthase significantly reduced L-carnitine-induced PGI2 and L-carnitine's protective effect. Furthermore, blockade the potential PGI2 receptors, including PGI2 receptors (IP receptors), and peroxisome proliferator-activated receptors alpha and delta (PPARα and PPARδ), revealed that the siRNA-mediated blockage of PPARα considerably reduced the anti-apoptotic effect of L-carnitine. Conclusions: These findings suggest that L-carnitine protects cardiomyocytes from doxorubicin-induced apoptosis in part through PGI2 and PPARα-signaling pathways, which may potentially protect the heart from the severe toxicity of doxorubicin.",
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author = "Chao, {Hung Hsin} and Liu, {Ju Chi} and Hong, {Hong Jye} and Lin, {Jia Wei} and Chen, {Cheng Hsien} and Tzu-Hurng Cheng",
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T1 - L-carnitine reduces doxorubicin-induced apoptosis through a prostacyclin-mediated pathway in neonatal rat cardiomyocytes

AU - Chao, Hung Hsin

AU - Liu, Ju Chi

AU - Hong, Hong Jye

AU - Lin, Jia Wei

AU - Chen, Cheng Hsien

AU - Cheng, Tzu-Hurng

PY - 2011/1/21

Y1 - 2011/1/21

N2 - Background: Clinical use of doxorubicin is greatly limited by its severe cardiotoxic side effects. L-carnitine is a vitamin-like substance which has been successfully used in many cardiomyopathies, however, the intracellular mechanism(s) remain unclear. The objective of this study was set to evaluate the protective effect of L-carnitine on doxorubicin-induced cardiomyocyte apoptosis, and to explore its intracellular mechanism(s). Methods: Primary cultured neonatal rat cardiomyocytes were treated with doxorubicin (1 μM) with or without pretreatment with L-carnitine (1-30 mM). Lactate dehydrogenase assay, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling staining, and flow cytometry measurement were used to assess cytotoxicity and apoptosis. Fluorescent probes 2',7'-dichlorofluorescein diacetate and chemiluminescence assay of superoxide production were used to detect the production of reactive oxygen species. Western blotting was used to evaluate the quantity of cleaved caspase-3, cytosol cytochrome c, and Bcl-x L expression. Results: L-carnitine inhibited doxorubicin-induced reactive oxygen species generation and NADPH oxidase activation, reduced the quantity of cleaved caspase-3 and cytosol cytochrome c, and increased Bcl-x L expression, resulting in protecting cardiomyocytes from doxorubicin-induced apoptosis. In addition, L-carnitine was found to increase the prostacyclin (PGI2) generation in cardiomyocytes. The siRNA transfection for PGI2 synthase significantly reduced L-carnitine-induced PGI2 and L-carnitine's protective effect. Furthermore, blockade the potential PGI2 receptors, including PGI2 receptors (IP receptors), and peroxisome proliferator-activated receptors alpha and delta (PPARα and PPARδ), revealed that the siRNA-mediated blockage of PPARα considerably reduced the anti-apoptotic effect of L-carnitine. Conclusions: These findings suggest that L-carnitine protects cardiomyocytes from doxorubicin-induced apoptosis in part through PGI2 and PPARα-signaling pathways, which may potentially protect the heart from the severe toxicity of doxorubicin.

AB - Background: Clinical use of doxorubicin is greatly limited by its severe cardiotoxic side effects. L-carnitine is a vitamin-like substance which has been successfully used in many cardiomyopathies, however, the intracellular mechanism(s) remain unclear. The objective of this study was set to evaluate the protective effect of L-carnitine on doxorubicin-induced cardiomyocyte apoptosis, and to explore its intracellular mechanism(s). Methods: Primary cultured neonatal rat cardiomyocytes were treated with doxorubicin (1 μM) with or without pretreatment with L-carnitine (1-30 mM). Lactate dehydrogenase assay, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling staining, and flow cytometry measurement were used to assess cytotoxicity and apoptosis. Fluorescent probes 2',7'-dichlorofluorescein diacetate and chemiluminescence assay of superoxide production were used to detect the production of reactive oxygen species. Western blotting was used to evaluate the quantity of cleaved caspase-3, cytosol cytochrome c, and Bcl-x L expression. Results: L-carnitine inhibited doxorubicin-induced reactive oxygen species generation and NADPH oxidase activation, reduced the quantity of cleaved caspase-3 and cytosol cytochrome c, and increased Bcl-x L expression, resulting in protecting cardiomyocytes from doxorubicin-induced apoptosis. In addition, L-carnitine was found to increase the prostacyclin (PGI2) generation in cardiomyocytes. The siRNA transfection for PGI2 synthase significantly reduced L-carnitine-induced PGI2 and L-carnitine's protective effect. Furthermore, blockade the potential PGI2 receptors, including PGI2 receptors (IP receptors), and peroxisome proliferator-activated receptors alpha and delta (PPARα and PPARδ), revealed that the siRNA-mediated blockage of PPARα considerably reduced the anti-apoptotic effect of L-carnitine. Conclusions: These findings suggest that L-carnitine protects cardiomyocytes from doxorubicin-induced apoptosis in part through PGI2 and PPARα-signaling pathways, which may potentially protect the heart from the severe toxicity of doxorubicin.

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