scholarly journals Sodium nitroprusside induces cell death and cytoskeleton degradation in adult rat cardiomyocytes in vitro: implications for anthracycline-induced cardiotoxicity

2012 ◽  
Vol 56 (2) ◽  
pp. 15 ◽  
Author(s):  
M. Chiusa ◽  
F. Timolati ◽  
J.C. Perriard ◽  
T.M. Suter ◽  
C. Zuppinger
Keyword(s):  
Cell Death ◽  
Sodium Nitroprusside ◽  
Rat Cardiomyocytes ◽  
Adult Rat

Abstract 165: The 18 kDa FGF-2 Prevents the Doxorubicin-induced Cardiac Damage and Amp-activated Kinase Activation, in vitro and in vivo

2015 ◽  
Vol 117 (suppl_1) ◽  
Author(s):  
Navid Koleini ◽  
Jon Jon Santiago ◽  
Barbara E Nickel ◽  
Robert Fandrich ◽  
Davinder S Jassal ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Cell Death ◽  
Left Ventricular ◽  
Left Ventricular Ejection ◽  
Wild Type ◽  
Rat Cardiomyocytes ◽  
Ventricular Ejection Fraction ◽  
Compound C

Introduction: Protection of the heart from chemotherapeutic (Doxorubicin, DOX) drug-induced toxicity is a desirable goal, to limit side effects of cancer treatments. DOX toxicity has been linked to the activation (phosphorylation) of the AMP-activated kinase, AMPK. The 18 kDa low molecular weight isoform of fibroblast growth factor 2 (Lo-FGF-2) is a known cardioprotective and cytoprotective agent. In this study we have tested the ability of Lo-FGF-2 to protect from DOX-induced damage in rat cardiomyocytes in vitro, and in transgenic mouse models in vivo, in relation to AMPK activation. Methods: Rat neonatal cardiomyocytes in culture were exposed to DOX (0.5 μM) in the presence or absence of pre-treatment Lo-FGF-2 (10 ng/ml). Compound C was used to block phosphorylation (activity) of AMPK. Levels of cell viability/death (using Calcein-AM/Propidium iodide assay), phospho -and total AMPK, and apoptotic markers such as active caspase 3 were analyzed. In addition, transgenic mice expressing only Lo-FGF2, and wild type mice, expressing both high molecular weight (Hi-FGF2) as well as Lo-FGF2 were subjected to DOX injection (20 mg/kg, intraperitoneal); echocardiography was used to examine cardiac function at baseline and at 10 days post-DOX. Results: DOX-induced cell death of cardiomyocytes in culture was maximal at 24 hours post-DOX coinciding with significantly increased in activated (phosphorylated) AMPK. Compound C attenuated DOX-induced cardiomyocyte loss. Pre-incubation with Lo-FGF-2 decreased DOX induced cell death, and also attenuated the phosphorylation of AMPK post-DOX. Relative levels of phospho-AMPK were lower in the hearts of Lo-FGF2-expressing male mice compared to wild type. DOX-induced loss of contractile function (left ventricular ejection fraction and endocardial velocity) was negligible in Lo-FGF2-expressing mice but significant in wild type mice. Conclusion: Lo-FGF-2 protects the heart from DOX-induced damage in vitro and in vivo, by a mechanism likely involving an attenuation of AMPK activity.

Adrenergic activation of glycogen phosphorylase in primary culture diabetic cardiomyocytes

1992 ◽  
Vol 262 (3) ◽  
pp. H649-H653 ◽  
Author(s):  
J. A. Buczek-Thomas ◽  
S. R. Jaspers ◽  
T. B. Miller
Keyword(s):  
Hypersensitive Response ◽  
Adrenergic Receptor ◽  
Glycogen Phosphorylase ◽  
Receptor Activation ◽  
Receptor Stimulation ◽  
Beta Adrenergic ◽  
Rat Cardiomyocytes ◽  
Adult Rat ◽  
Adrenergic Activation

The basis of catecholamine-induced activation of glycogen phosphorylase was investigated in adult rat cardiomyocytes isolated from normal and alloxan-diabetic animals. Cells derived from diabetic animals exhibited a hypersensitive response to epinephrine stimulation that was apparent 3 h after cell isolation and was further enhanced on maintenance of the myocytes in culture for 24 h. Normal cells initially lacked the hypersensitive response to epinephrine stimulation, although on maintenance of these cells in culture for 24 h, the hypersensitive response was acquired in vitro. To assess alpha- and beta-adrenergic mediation of the response, normal and diabetic cardiomyocytes were incubated with propranolol, a beta-blocker, before direct alpha 1-receptor stimulation with phenylephrine. Both normal and diabetic myocytes failed to undergo activation of phosphorylase in 3- or 24-h cell cultures. In addition, the effects of epinephrine on phosphorylase activation were completely inhibited by propranolol, whereas prazosin, an alpha-blocker, was unsuccessful. The present data suggest that the hypersensitive response of glycogen phosphorylase in normal and diabetic cardiomyocytes is solely mediated through beta-adrenergic receptor activation.

Evidence for functional sodium and calcium ion channels in the membrane of cultured cardiomyocytes of the adult rat

1983 ◽  
Vol 61 (11) ◽  
pp. 1312-1316 ◽  
Author(s):  
S. L. Jacobson ◽  
C. B. Kennedy ◽  
G. A. R. Mealing
Keyword(s):  
Action Potentials ◽  
Rat Cardiomyocytes ◽  
Adult Rat ◽  
Culture Cells ◽  
Calcium Ion Channels ◽  
Spontaneous Action ◽  
Fetal Bovine ◽  
Spontaneous Action Potentials

Characteristics are reported for electrical activity of adult rat cardiomyocytes in long-term primary culture. Cells in vitro for 12 to 28 days have mean membrane potential of −53 mV, are electrically excitable, and some are spontaneously contractile. The action potential of these cells has a slow rate of depolarization and is abolished by methoxyverapamil (D-600) but not by tetrodotoxin (TTX). When cells are hyperpolarized by passage of an inward current, spontaneous action potentials cease and action potentials evoked by depolarizing pulses are then TTX sensitive. Fetal bovine serum is a constituent of the culture medium. Its temporary removal causes spontaneous contractility to cease but the cells remain electrically excitable.

scholarly journals Cardiac-targeted PIASy Gene Silencing Mediates deSUMOylation of Caveolin-3 and Prevents Ischemia/reperfusion-induced Nav1.5 Down-regulation and Ventricular Arrhythmias

2021 ◽  
Author(s):  
Chenchen Hu ◽  
Xin Wei ◽  
Jinmin Liu ◽  
Linlin Han ◽  
Chengkun Xia ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Ventricular Arrhythmias ◽  
Heart Diseases ◽  
Ischemia Reperfusion ◽  
Intraventricular Injection ◽  
Membrane Expression ◽  
Down Regulation ◽  
Rat Cardiomyocytes ◽  
Adult Rat

Abstract Background: Abnormal myocardial expression and function of Nav1.5 causes lethal ventricular arrhythmias during myocardial ischemia-reperfusion (I/R). PIASy mediated Caveolin-3 (Cav-3) SUMO modification affects Cav-3 binding to ligand Nav1.5. PIASy activity is increased after myocardial I/R, whether or not this may be attributable to plasma membrane Nav1.5 downregulation and ventricular arrhythmias remains unclear. Methods: Using recombinant adeno-associated virus subtype 9 (AAV9), rat cardiac PIASy was silenced by intraventricular injection of PIASy shRNA. Two weeks later, the hearts were subjected to I/R, and electrocardiography was performed to assess malignant arrhythmias. Tissues from peri-infarct areas of the left ventricle were collected for molecular biological measurement. Results: We found that PIASy was upregulated by I/R, with increased SUMO2/3 modification of Cav-3, reduced membrane Nav1.5 density, and increased ventricular arrhythmia frequency. These effects were significantly reversed by PIASy silencing. In addition, PIASy silencing enhanced Cav-3 binding to Nav1.5 and prevented I/R-induced Nav1.5 re-localization. Using in vitro models of HEK293T cells and isolated adult rat cardiomyocytes exposed to hypoxia/reoxygenation (H/R), this reserch further confirmed that PIASy promoted Cav-3 modification by SUMO2/3 and Nav1.5/Cav-3 dissociation after H/R. Mutation of the SUMO Consensus Sites Lysine in Cav-3 (K38R or K144R) alters the membrane expression levels of Nav1.5 and Cav-3 before and after H/R in HEK293T cells. Conclusions: I/R-induced cardiac PIASy activation contributes to Cav-3 SUMOylation by SUMO2/3 and dysregulated Nav1.5- related ventricular arrhythmias. Cardiac-targeted PIASy gene silencing mediates deSUMOylation of Cav-3 and prevents I/R-induced Nav1.5 down-regulation and ventricular arrhythmias in rats, identifying PIASy as a potential therapeutic target for relevant life-threatening arrhythmias in patients with ischemic heart diseases.

Abstract 14664: SIRT3 Protects Cardiomyocytes From Doxorubicin-induced Mitochondrial Damage and Cell-death by Activating Opa1

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Sadhana Samant ◽  
Vinodkumar Pillai ◽  
Don Wolfgeher ◽  
Mahesh Gupta
Keyword(s):  
Cell Death ◽  
Mitochondrial Damage ◽  
Confocal Imaging ◽  
Neonatal Rat ◽  
Cancer Drug ◽  
Gtpase Activity ◽  
Rat Cardiomyocytes ◽  
Apoptotic Markers ◽  
Mitochondrial Functions

Introduction: The anti-cancer drug therapy with doxorubicin (Dox) is known to produce cardiomyopathy associated with fragmentation of mitochondria and excess production of reactive oxygen species (ROS). Sirtuin3 (SIRT3), a mitochondrial deacetylase regulates many mitochondrial functions including energy production, ROS synthesis and cell-death. A role of SIRT3 in regulating the aging process and aging-associated diseases like heart failure, cancer and diabetes has been also documented. This study was undertaken to investigate whether SIRT3 can protect cardiomyocytes from Dox-induced mitochondrial damage and cell-death. Methods and Results: Neonatal rat cardiomyocytes were over expressed with a vector synthesizing SIRT3 or a mock vector. Cells were treated with Dox (100nM) for 72 hours and cell-death was analyzed by measuring TUNEL-positive nuclei, ROS synthesis and expression of apoptotic markers by the western blotting. There was marked reduction of TUNEL-positive nuclei, ROS levels and expression of apoptotic markers such as Bax, Bim and FasL in SIRT3 over expressing cells, compared to control cells received the mock vector. Confocal imaging of myocytes revealed that Dox-treatment caused swelling and fragmentation of mitochondria in control cells, but not in cells over expressed with SIRT3. To understand the mechanism involved in SIRT3-mediated protection of mitochondria we analyzed acetylation of proteins involved in regulating mitochondrial fusion-fission dynamics. We found that OPA1 (optic atrophy-1), an essential inner membrane fusion protein, was hyper-acetylated in Dox-treated cells, leading to its reduced GTPase activity. SIRT3 was capable of deacetylating and augmenting GTPase activity of OPA1 both in vitro and in vivo assay conditions. Furthermore, by studying OPA1 null cells we found that it is necessary for SIRT3-mediated protection of cardiomyocytes from mitochondrial damage and cell-death. Conclusions: These results indicated that SIRT3-dependent activation of OPA1 contributes to preservation of mitochondrial population and protection of cardiomyocytes from Dox-mediated cell-death. We believe that SIRT3 could be utilized as a therapeutic target for treatment of Dox-induced cardiomyopathy of the heart.

P3116New role of EPAC1 in Anthracycline-induced cardiotoxicity and anticancer therapy

2019 ◽  
Vol 40 (Supplement_1) ◽  
Author(s):  
A Belhadef ◽  
M Ribeiro ◽  
M Mazevet ◽  
M Laudette ◽  
B Crozatier ◽  
...  
Keyword(s):  
Cell Death ◽  
Mitochondrial Membrane ◽  
Reactive Oxygen Species Generation ◽  
Neonatal Rat ◽  
Dna Intercalation ◽  
Mice Model ◽  
Rat Cardiomyocytes ◽  
Knock Out

Abstract Introduction Doxorubicin (Dox) is an anthracycline commonly used to treat many types of cancer; unfortunately this chemotherapeutic agent induces many side effects such as cardiotoxicity leading to dilated cardiomyopathy (DCM). The cardiotoxicity of Dox has been related to reactive oxygen species generation, DNA intercalation, topoisomerase II inhibition and bioenergetics alterations leading to cardiomyocyte death. Objective Nowadays the challenge is to find new treatment options aiming at reducing Dox cardiotoxicity. Epac (exchange protein directly activated by cAMP) signaling could be worth investigating as Epac activates small G proteins which are known to be involved in Dox-induced cardiotoxicity. Methods We investigated the time/dose-dependent Dox effect on Epac signaling in both in vivo mice model (C57Bl63/ Knock-out Epac1 mice, iv injections, 12mg/kg cumulative dose) and in vitro (primary culture of neonatal rat cardiomyocytes (NRVM, 24h, Dox 1μM). Results In vivo, Dox-treated mice developed a DCM associated with Ca2+ homeostasis dysfunction (increase of Ca2+ waves and Ca2+ leaks). In vitro, as measured by flow cytometry and western blot, Dox (1μM) induced DNA damages and cell death in NRVM. This cell death is associated with apoptotic features including mitochondrial membrane permeabilization, caspase activation and cell size reduction. The inhibition of Epac1 (ESI09, CE3F4) decreased Dox-induced DNA damage, loss of mitochondrial membrane potential, apoptosis and finally cardiomyocyte death. Moreover, in vivo, Epac1 KO mice were protected against Dox-induced cardiotoxicity by unaltered cardiac function (no DCM) and calcium homeostasis at 15 weeks post-treatment. Conclusion Inhibition of Epac1 could be a valuable therapeutic strategy to limit Dox-induced cardiomyopathy during cancer chemotherapy. Indeed, preliminary data show also that preventing Dox-induced cardiotoxicity, the inhibition of Epac1 can also potentiate cancerous cells death. Acknowledgement/Funding Labex Lermit (ANR 0033), Torino and Inserm

scholarly journals The Nitrone Disodium 2,4-Sulphophenyl-N-Tert-Butylnitrone is Without Cytoprotective Effect on Sodium Nitroprusside-Induced Cell Death in N1E-115 Neuroblastoma Cells in vitro

2007 ◽  
Vol 28 (1) ◽  
pp. 24-28 ◽  
Author(s):  
Atticus H Hainsworth ◽  
Nasrin Bhuiyan ◽  
A Richard Green
Keyword(s):  
Cell Death ◽  
Free Radical ◽  
Sodium Nitroprusside ◽  
Neurovascular Unit ◽  
Neuroblastoma Cells ◽  
Mitochondrial Complex ◽  
Complex Ii ◽  
Radical Trapping

Disodium 2,4-sulphophenyl- N-tert-butylnitrone (NXY-059) is a novel free radical-trapping compound that is neuroprotective in both rodent and primate models of acute ischaemic stroke. Neuroprotection in vitro by NXY-059 has not been reported previously, and we have now investigated whether such an effect can be detected using a simple cell culture model of neurotoxicity. Neuron-like cells of the neuroblastoma-derived clonal cell line N1E-115 were exposed to the free radical-generating agent sodium nitroprusside (SNP), which produced a concentration-dependent reduction in mitochondrial complex II activity 24 h later (EC50 approximately 100 micromolar). Cell death induced by SNP (100 micromolar), assessed either by an increased proportion of apoptotic nuclear morphology or by mitochondrial complex II activity, was inhibited by a cocktail of known antioxidants (ascorbate, reduced glutathione, and dithiothreitol, all at 100 micromolar) but not by NXY-059 at a concentration known to be neuroprotective in vivo (300 micromolar). Disodium 2,4-sulphophenyl- N-tert-butylnitrone was also without effect on H2O2-mediated cytotoxicity. These results support recent data suggesting that in vivo NXY-059 probably acts at the neurovascular unit rather than at an intracellular site as a neuroprotective agent.

Daily administration of interleukin-18 causes myocardial dysfunction in healthy mice

2005 ◽  
Vol 289 (2) ◽  
pp. H708-H714 ◽  
Author(s):  
Per Reidar Woldbæk ◽  
Jørn Bodvar Sande ◽  
Tævje Andreas Strømme ◽  
Per Kristian Lunde ◽  
Srdjan Djurovic ◽  
...  
Keyword(s):  
Myocardial Hypertrophy ◽  
Diastolic Pressure ◽  
Myocardial Dysfunction ◽  
Fold Increase ◽  
Left Ventricular ◽  
Daily Administration ◽  
Lv Function ◽  
Rat Cardiomyocytes ◽  
Adult Rat

Although increased levels of circulating interleukin (IL)-18 have been demonstrated in patients with cardiovascular diseases, the functional consequences of chronically increased circulating IL-18 with respect to myocardial function have not been defined. Thus we aimed to examine the effects of chronic IL-18 exposure on left ventricular (LV) function in healthy mice. Moreover, to clarify whether IL-18 has direct effects on the cardiomyocyte, we examined effects of IL-18 on cardiomyocytes in vitro. After 7 days of daily intraperitoneal injections of 0.5 μg IL-18 in healthy mice, a 40% ( P < 0.05) reduction in the LV maximal positive derivative, a 25% ( P < 0.05) reduction in the LV maximal rate of pressure decay, and a 2.8-fold ( P < 0.001) increase in the LV end-diastolic pressure were measured, consistent with myocardial dysfunction. Furthermore, we measured a 75% ( P < 0.05) reduction in β-adrenergic responsiveness to isoproterenol. IL-18 induced myocardial hypertrophy, and there was a 2.9-fold increase ( P < 0.05) in atrial natriuretic peptide mRNA expression in the LV myocardium. In vitro examinations of isolated adult rat cardiomyocytes being stimulated with IL-18 (0.1 μg/ml) exhibited an increase in peak Ca2+ transients ( P < 0.05) and in diastolic Ca2+ concentrations ( P < 0.05). In conclusion, this study shows that daily administration of IL-18 in healthy mice causes LV myocardial dysfunction and blunted β-adrenergic responsiveness to isoproterenol. A direct effect of IL-18 on the cardiomyocyte in vitro was demonstrated, suggesting that IL-18 reduces the responsiveness of the myofilaments to Ca2+. Finally, induction of myocardial hypertrophy by IL-18 indicates a role for this cytokine in myocardial remodeling.

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