Doxorubicin Induces Apoptosis by Activation of Caspase-3 in Cultured Cardiomyocytes In Vitro and Rat Cardiac Ventricles In Vivo

Ueno, Michihiko; Kakinuma, Yoshihiko; Yuhki, Koh-ichi; Murakoshi, Nobuyuki; Iemitsu, Motoyuki; Miyauchi, Takashi; Yamaguchi, Iwao

doi:10.1254/jphs.fp0050980

Cited by 163 publications

(125 citation statements)

References 31 publications

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“…It is pertinent to speculate that the same effects could be attained in vivo using a subchronic or chronic treatment with lower, pharmacological doses due to tissue drug accumulation. In fact, most alterations induced by Dox on H9c2 myoblasts, such as mitochondrial and nuclear damage, cytoplasm vacuolization, alterations in sarcomeric proteins, and apoptosis, are indeed observed in cardiac tissue after in vivo Dox treatment (Grimmond and Beerman 1982;Iwasaki and Suzuki 1991;Jang et al 2004;Jones et al 1990;Ueno et al 2006;Unverferth et al 1981;Villani et al 1990). …”

Section: Discussionmentioning

confidence: 99%

“…Caspase 9 is control relatively to healthy (asterisk) and apoptotic cells dependent on the mitochondrial pathway for apoptosis (Garrido et al 2006;Schafer and Kornbluth 2006), confirming that mitochondria are involved in Doxinduced toxicity on H9c2 myoblasts. In fact, both mitochondrial-dependent (Ueno et al 2006) and mitochondrial-independent (Jang et al 2004) pathways have been described as involved in in vivo apoptosis induced by Dox in cardiac cells. LDH release assays also confirmed that Dox causes cell death, although the method itself measures necrosis, which can occur in vitro sequentially to apoptosis induction or occur as a primary event.…”

Section: Discussionmentioning

confidence: 99%

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Morphological alterations induced by doxorubicin on H9c2 myoblasts: nuclear, mitochondrial, and cytoskeletal targets

et al. 2008

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Doxorubicin (Dox) is a very potent antineoplastic agent used against several types of cancer, despite a cumulative cardiomyopathy that reduces the therapeutic index for treatment. H9c2 myoblast cells have been used as an in vitro model to study biochemical alterations induced by Dox treatment on cardiomyocyte cells. Despite the extensive work already published, few data are available regarding morphological alterations of H9c2 cells during Dox treatment. The purpose of the present work was to evaluate Doxinduced morphological alterations in H9c2 myoblasts, focusing especially on the nuclei, mitochondria, and structural fibrous proteins. Treatment of H9c2 cell with low concentrations of Dox causes alterations in fibrous structural proteins including the nuclear lamina and sarcomeric cardiac myosin, as well as mitochondrial depolarization and fragmentation, membrane blebbing with cell shape changes, and phosphatidylserine externalization. For higher Dox concentrations, more profound alterations are evident, including nuclear swelling with disruption of nuclear membrane structure, mitochondrial swelling, and extensive cytoplasm vacuolization. The results obtained indicate that Dox causes morphological alterations in mitochondrial, nuclear, and fibrous protein structures in H9c2 cells, which are dependent on the drug concentration. Data obtained with the present study allow for a better characterization of the effects of Dox on H9c2 myoblasts, used as a model to study Dox-induced cardiotoxicity. The results obtained also provide new and previously unknown targets that can contribute to understand the mechanisms involved in the cardiotoxicity of Dox.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Morphological alterations induced by doxorubicin on H9c2 myoblasts: nuclear, mitochondrial, and cytoskeletal targets

et al. 2008

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“…Cells were stimulated with ADR, 3 g/ml (5 M) for 3 d. 42,43 Supernatants were retrieved for AGE ELISA. In other studies, podocytes were stimulated with S-100B.…”

Section: Cell Culturementioning

confidence: 99%

RAGE Mediates Podocyte Injury in Adriamycin-induced Glomerulosclerosis

Jiang¹,

Ananthakrishnan²,

Qu³

et al. 2008

Journal of the American Society of Nephrology

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In the kidney, the receptor for advanced glycation end products (RAGE) is principally expressed in the podocyte at low levels, but is upregulated in both human and mouse glomerular diseases. Because podocyte injury is central to proteinuric states, such as the nephrotic syndrome, the murine adriamycin nephrosis model was used to explore the role of RAGE in podocyte damage. In this model, administration of the anthracycline antibiotic adriamycin provokes severe podocyte stress and glomerulosclerosis. In contrast to wild-type animals, adriamycin-treated RAGE-null mice were significantly protected from effacement of the podocyte foot processes, albuminuria, and glomerulosclerosis. Administration of adriamycin induced rapid generation of RAGE ligands, and treatment with soluble RAGE protected against podocyte injury and glomerulosclerosis. In vitro, incubation of RAGE-expressing murine podocytes with adriamycin stimulated AGE formation, and treatment with RAGE ligands rapidly activated nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase, via p44/p42 MAP kinase signaling, and upregulated pro-fibrotic growth factors. These data suggest that RAGE may contribute to the pathogenesis of podocyte injury in sclerosing glomerulopathies such as focal segmental glomerulosclerosis.

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“…Apoptosis has been identified in various cardiac disease states, for example, in ischemia and reperfusion injury (Krijnen et al 2002;Eefting et al 2004;Matsushita et al 2005;Liang et al 2006) or doxorubicin toxicity (Ueno et al 2006). Evans blue staining of myocytes has been associated with apoptosis in muscular dystrophy (Matsuda, 1995).…”

Section: Introductionmentioning

confidence: 99%

Evans Blue Staining of Cardiomyocytes Induced by Myocardial Contrast Echocardiography in Rats: Evidence for Necrosis Instead of Apoptosis

Miller

Peng

Dou

et al. 2007

Ultrasound in Medicine & Biology

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High Mechanical Index (MI) echocardiography with contrast agent has been shown to induce Evans blue staining of cardiomyocytes, seen one day after exposure, in addition to contraction band necrosis, seen immediately after exposure. This research examined the roles of necrosis vs. apoptosis in these bioeffects. Myocardial contrast echocardiography at high MI with 1:4 ECG triggering was performed in anesthetized rats at 1.5 MHz. Histologically observable cell injury was accumulated by infusing a high dose of 50 μl/kg Definity® via tail vein for 5 min at the start of 10 min of scanning. Evans blue dye or propidium iodide was injected as an indicator of cardiomyocyte plasma membrane integrity. Histological sections were stained using the TUNEL method for labeling nuclei with DNA degradation (e.g. apoptosis). Evans blue fluorescent cells were counted on frozen sections or on hematoxylin-stained and TUNEL labeled paraffin sections. In addition, transmission electron microscopy was used to assess potential apoptotic nuclei. Hypercontraction and propidium iodide staining were observed immediately after imaging-exposure. Although TUNEL positive cells were evident after 4 h, these also had indications of contraction band necrosis and features of apoptosis were not confirmed by electron microscopy. Inflammatory cell infiltration was evident after 24 h. A second, more subtle injury was recognized by Evans blue staining, with minimal inflammatory cell infiltration at the morphologically intact stained cells after 24 h. Apoptosis was not detected by the TUNEL method in the cardiomyocytes stained with Evans blue at 24 h. However, Evans blue stained cell numbers declined after 48 h, with continued inflammatory cell infiltration. The initial insult for Evans blue stained cardiomyocytes apparently induced partial permeability of the plasma membrane, which led to gradual degeneration (but not apoptosis) and necrosis after 24-48 h.

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Doxorubicin Induces Apoptosis by Activation of Caspase-3 in Cultured Cardiomyocytes In Vitro and Rat Cardiac Ventricles In Vivo

Cited by 163 publications

References 31 publications

Morphological alterations induced by doxorubicin on H9c2 myoblasts: nuclear, mitochondrial, and cytoskeletal targets

Morphological alterations induced by doxorubicin on H9c2 myoblasts: nuclear, mitochondrial, and cytoskeletal targets

RAGE Mediates Podocyte Injury in Adriamycin-induced Glomerulosclerosis

Evans Blue Staining of Cardiomyocytes Induced by Myocardial Contrast Echocardiography in Rats: Evidence for Necrosis Instead of Apoptosis

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