Proteomic investigation of embryonic rat heart-derived H9c2 cell line sheds new light on the molecular phenotype of the popular cell model

Moraea pallida Bak. (yellow tulp) poisoning is the most important cardiac glycoside-induced intoxication in ruminants in South Africa. The toxic principle, 1α, 2α-epoxyscillirosidine, is a bufadienolide. To replace the use of sentient animals in toxicity testing, the aim of this study was to evaluate the cytotoxic effects of epoxyscillirosidine on rat embryonic cardiomyocytes (H9c2 cell line). This in vitro cell model can then be used in future toxin neutralization or toxico-therapy studies. Cell viability, evaluated with the methyl blue thiazol tetrazolium (MTT) assay, indicated a hormetic dose/concentration response, characterized by a biphasic low dose stimulation and high dose inhibition. Increased cell membrane permeability and leakage, as expected with necrotic cells, were demonstrated with the lactate dehydrogenase (LDH) assay. The LC50 was 382.68, 132.28 and 289.23 μM for 24, 48, and 72 h respectively. Numerous cytoplasmic vacuoles, karyolysis and damage to the cell membrane, indicative of necrosis, were observed at higher doses. Ultra-structural changes suggested that the cause of H9c2 cell death, subsequent to epoxyscillirosidine exposure, is necrosis, which is consistent with myocardial necrosis observed at necropsy. Based on the toxicity observed, and supported by ultra-structural findings, the H9c2 cell line could be a suitable in vitro model to evaluate epoxyscillirosidine neutralization or other therapeutic interventions in the future.

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Regulation of β-adrenoceptor density and mRNA levels in the rat heart cell-line H9c2

Biochemical Journal ◽

10.1042/bj3170925 ◽

1996 ◽

Vol 317 (3) ◽

pp. 925-931 ◽

Cited By ~ 42

Author(s):

Volker DANGEL ◽

Jeanette GIRAY ◽

Dieter RATGE ◽

Hermann WISSER

Keyword(s):

Cell Line ◽

Half Life ◽

Rat Heart ◽

Radioligand Binding ◽

Mrna Level ◽

Fold Increase ◽

Heart Cell ◽

H9c2 Cell ◽

Heart Cells ◽

Mrna Levels

The regulation of the expression of β-adrenoceptors (β-ARs) is not thoroughly understood. We demonstrate that the rat heart cell-line H9c2 expresses both β1- and β2-ARs. In radioligand-binding experiments, the maximal binding capacity of (-)-[125I]-iodocyanopindolol was determined as 18±0.6 fmol/mg of protein with a KD of 35.4±4.1 pM. Competitive radioligand-binding experiments with subtype-specific β-antagonists reveal a subtype ratio of β1- to β2-ARs of 29%:71%. With competitive reverse-transcriptase PCR we found β2-mRNA to be up to 1600 times more frequent than β1-mRNA. Treatment of the H9c2 cell-line with the β-adrenergic agonist (-)-isoproterenol (10-6 M), the antagonist (-)-propranolol (10-6 M) and the glucocorticoid dexamethasone (500 nM) induces regulatory effects on both the β-AR protein and mRNA level. Isoproterenol treatment leads to down-regulation of the total receptor number by 56±4%, due to a decrease in β2-ARs, while maintaining the β1-AR number constant. On the transcription level, both β1-and β2-mRNAs are decreased by 30% and 42% respectively. mRNA stability measurements reveal a reduced half-life of β2-mRNA from 9.3 h to 6.5 h after isoproterenol treatment. Incubation of cells with (-)-propranolol does not affect the amounts of β-ARs and their mRNAs. Dexamethasone induces a 1.8±0.2-fold increase in β-AR number over the basal level as well as a 1.9±0.2-fold increase in the amount of β2-mRNA. Because the half-life of β2-mRNA was unaffected by dexamethasone, the increased β2-mRNA level must be due to an enhanced transcription rate. The β1-mRNA levels are unchanged during dexamethasone-incubation of the cells. Our data clearly demonstrate that treatment of H9c2 rat heart cells with isoproterenol and dexamethasone induces alterations in the level of RNA stability as well as gene transcription, leading to altered receptor numbers.

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Over‐expression of FXRG and miR‐1 increases formation of RISC complexes in H9C2 cell line

The FASEB Journal ◽

10.1096/fasebj.27.1_supplement.1189.4 ◽

2013 ◽

Vol 27 (S1) ◽

Author(s):

Lisa Walker ◽

Karen Dockstader ◽

Dobromir Slavov ◽

Carmen Sucharov

Keyword(s):

Cell Line ◽

H9c2 Cell ◽

Over Expression ◽

H9c2 Cell Line

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THE VIABILITY TEST OF SAPPAN WOOD (CAESALPINIA SAPPAN L.) ETHANOL EXTRACT IN THE H9C2 CELL LINE

International Journal of Applied Pharmaceutics ◽

10.22159/ijap.2020.v12s3.39479 ◽

2020 ◽

pp. 76-78

Author(s):

IFA SULISTIYORINI ◽

RATU SAFITRI ◽

RONNY LESMANA ◽

MAS RIZKY A. A. SYAMSUNARNO

Keyword(s):

Cell Viability ◽

Cell Line ◽

Control Cell ◽

Ethanol Extract ◽

Optimal Dose ◽

H9c2 Cell ◽

Caesalpinia Sappan ◽

Viable Cells ◽

Dose Concentration ◽

H9c2 Cell Line

Objective: In this study, the embryonic rat cardiomyocyte cell line H9C2 was used to investigate the cardiotoxicity effect of sappan wood ethanol extract (SWEE). Methods: Sappan wood was extracted in 96% ethanol and divided into dose concentrations of 2.5, 5, 10, 50, 100, and 150 μg/ml, with deferiprone used as a control. Cell viability was assessed using the PrestoBlue Cell Viability Reagent, according to manufacturer protocols. Results: Microscopic examination showed that the cell viability of H9C2 was preserved by SWEE treatments at a dose of 10 μg/ml and suggested dose concentrations of 50 μg/ml of SWEE. The percentage of viable cells was greater than 95% with a dose concentration of 10 μg/ml of SWEE, but it was significantly reduced with a dose concentration of 50 μg/ml of SWEE (p<0.05). Conclusion: The optimal dose concentration of SWEE to reach 95% cell viability was 10 μg/ml.

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H9C2 cell line as a model of anthracycline cardiotoxicity

Journal of Molecular and Cellular Cardiology ◽

10.1016/s0022-2828(01)90260-3 ◽

2001 ◽

Vol 33 (6) ◽

pp. A66

Author(s):

Thomas J. L'Ecuyer ◽

Richard S. Vander Heide

Keyword(s):

Cell Line ◽

H9c2 Cell ◽

Anthracycline Cardiotoxicity ◽

H9c2 Cell Line

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Cytoprotective potential of anti-ischemic drugs against chemotherapy-induced cardiotoxicity in H9c2 myoblast cell line

Acta Pharmaceutica ◽

10.2478/acph-2013-0038 ◽

2013 ◽

Vol 63 (4) ◽

pp. 493-503 ◽

Cited By ~ 4

Author(s):

Tiam Feridooni ◽

Chris Mac Donald ◽

Di Shao ◽

Pollen Yeung ◽

Remigius U. Agu

Keyword(s):

Cell Death ◽

Cell Line ◽

Cancer Drug ◽

H9c2 Cell ◽

Drug Induced ◽

Cardiac Model ◽

Myoblast Cell Line ◽

Myoblast Cell ◽

H9c2 Cell Line

Abstract To investigate potential prevention or attenuation of anti- cancer drug induced cardiotoxicity using anti-ischemic drugs, a rat myoblast (H9c2) cell line was used as our in vitro cardiac model. Irinotecan and doxorubicin were found to be cytotoxic for the H9c2 cell line with IC50 of 30.69 ± 6.20 and 20.94 ± 6.05 mmol L-1, respectively. 5-Flurouracil and cladribine were not cytotoxic and thus IC50 could not be calculated. When 100 mmol L-1 doxorubicin was incubated for 72 hours with 50 mmol L-1 diltiazem, 100 mmol L-1 dexrazoxane and 100 mmol L-1 losartan, respectively, there was a 58.7 ± 10.2, 52.2 ± 11.7 and 44.7 ± 5.4 % reduction in cell death. When 200 mmol L-1 irinotecan was incubated for 72 hours with 100 mmol L-1 dexrazoxane, losartan and diltiazem, respectively, a 27.7 ± 6.9, 25.6 ± 5.1, and 19.1 ± 2.3 % reduction in cell death was observed. Our data suggests that losartan and diltiazem were as effective as dexrazoxane in protecting the cells against irinotecan- and doxorubicin-induced cell toxicity. These findings offer potential uses of anti- -ischemic drugs for ablation of cytotoxicity in response to mitochondrial injury, thereby improving patient outcomes and reducing health-care costs.

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