Journal: BMC complementary medicine and therapies

Article Title: Ginsenoside Rg3 attenuates myocardial ischemia/reperfusion-induced ferroptosis via the keap1/Nrf2/GPX4 signaling pathway.

doi: 10.1186/s12906-024-04492-4

Figure Lengend Snippet: Fig. 3 Erastin reversed the protective effect of ginsenoside Rg3 against OGD/R-induced injury in H9C2 cardiomyocytes. (A, B) Ferrous iron (Fe2+) and glutathione (GSH) levels in H9C2 cells were examined by kits. (C–E) The protein levels of GPX4 and FTH1 in H9C2 cells were examined by Western blot ting. The data are shown as the means ± SDs (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, the Ctrl group, the OGD/R + Rg3 group or the OGD/R + Rg3 + Erastin group vs. the OGD/R group

Article Snippet: Cell culture and oxygen-glucose deprivation/reperfusion (OGD/R) model H9C2 cardiomyocytes were obtained from BeiNa Biological Technology Co., Ltd. and cultured in DMEM supplemented with 10% FBS (Invitrogen) at 37 °C in a 5% CO2 incubator.

Techniques: Western Blot

Journal: Pharmacology

Article Title: Daphnetin Preconditioning Decreases Cardiac Injury and Susceptibility to Ventricular Arrhythmia following Ischaemia-Reperfusion through the TLR4/MyD88/NF-Κb Signalling Pathway

doi: 10.1159/000513631

Figure Lengend Snippet: Experimental protocol. a Different concentrations (2.5, 5, 10, and 20 mg/kg) of DAP and an equal volume of 0.9% saline were administered via daily gavage for 2 days before sham operation or I/R operation, and the myocardial infarct size and the myocardial enzyme levels were assessed 24 h after sham or I/R surgery. b DAP (10 mg/kg) and an equal volume of 0.9% saline were administered via daily gavage for 2 days before sham operation or I/R operation and for the long-term follow-up (treatment starting 24 h after sham or I/R operation). Electrophysiology, inflammation, apoptosis, oxidative stress, and protein expression were evaluated 24 h after sham or I/R surgery. Cardiac function was assessed before and 24 h, and 7 and 28 days after sham or I/R operation. c H9c2 cells were pretreated with different concentrations (5, 10, 20, and 40 μM) of DAP and 0.01% DMSO for 12 h and then subjected to 6 h of hypoxia and 6 h of reoxygenation or control treatment. Cell viability and apoptosis were tested in the H/R and control groups. d H9c2 cells were pretreated with DAP (20 μM) and 0.01% DMSO for 12 h and then subjected to 6 h of hypoxia and 6 h of reoxygenation or control treatment. Inflammation, oxidative stress, and protein expression were tested in the H/R and control groups. I/R, ischaemia/reperfusion; H/R, hypoxia/reoxygenation; DMSO, dimethylsulphoxide.

Article Snippet: Cell Culture and H/R Model Establishment H9c2 cells (iCell Bioscience Inc., Shanghai, China) were cultured in Dulbecco's modified Eagle's medium (DMEM; HyClone; GE Healthcare Life Sciences, Logan, UT, USA) supplemented with 10% foetal bovine serum at 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Saline, Expressing, Control

Journal: Pharmacology

Article Title: Daphnetin Preconditioning Decreases Cardiac Injury and Susceptibility to Ventricular Arrhythmia following Ischaemia-Reperfusion through the TLR4/MyD88/NF-Κb Signalling Pathway

doi: 10.1159/000513631

Figure Lengend Snippet: a Scatter dot plots quantifying the viability of H9c2 cells (n = 7). Representative flow cytometry (b) and scatter dot plots quantifying the apoptosis rate of H9c2 cells (c) (n = 7). d, e Scatter dot plots quantifying the mRNA levels of TNF-α and IL-6 in H9c2 cells (n = 6). f, g Scatter dot plots quantifying level of MDA and activity of SOD in H9c2 cells (n = 6). *p < 0.05 versus the control group; &p < 0.05 versus the H/R group; #p > 0.05 between the 20μM and 40 μM DAP treatment groups. DAP, daphnetin; MDA, malondialdehyde; SOD, superoxide dismutase; IL-6, interleukin-6; TNF-α, tumour necrosis factor-alpha; H/R, hypoxia/reoxygenation; FITC, fluorescein isothiocyanate.

Article Snippet: Cell Culture and H/R Model Establishment H9c2 cells (iCell Bioscience Inc., Shanghai, China) were cultured in Dulbecco's modified Eagle's medium (DMEM; HyClone; GE Healthcare Life Sciences, Logan, UT, USA) supplemented with 10% foetal bovine serum at 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Flow Cytometry, Activity Assay, Control

Journal: Pharmacology

Article Title: Daphnetin Preconditioning Decreases Cardiac Injury and Susceptibility to Ventricular Arrhythmia following Ischaemia-Reperfusion through the TLR4/MyD88/NF-Κb Signalling Pathway

doi: 10.1159/000513631

Figure Lengend Snippet: Representative Western blots of H9c2 cells (a) and scatter dot plots quantifying TLR4, MyD88, and p-NF-κB/P65 (b−d) (n = 6). *p < 0.05 versus the control group; &p < 0.05 versus the H/R group. H/R, hypoxia/reoxygenation.

Article Snippet: Cell Culture and H/R Model Establishment H9c2 cells (iCell Bioscience Inc., Shanghai, China) were cultured in Dulbecco's modified Eagle's medium (DMEM; HyClone; GE Healthcare Life Sciences, Logan, UT, USA) supplemented with 10% foetal bovine serum at 37°C in a humidified atmosphere containing 5% CO 2 .

Techniques: Western Blot, Control

Journal: BioMed Research International

Article Title: Interleukin-27 Protects Cardiomyocyte-Like H9c2 Cells against Metabolic Syndrome: Role of STAT3 Signaling

doi: 10.1155/2015/689614

Figure Lengend Snippet: Cytotoxic effects of high glucose, high sodium, or high cholesterol on the viability of H9c2 cells. (a) Cells were treated with glucose (25 mM, G), NaCl (250 mM, S), or cholesterol (300 μ g mL −1 , C), either alone or in different combinations, for 4, 8, 24, and 48 h. Cells were treated with IL-27 4 h before treatment with G, S, or C. Lactate dehydrogenase (LDH) release was measured as marker of cell injury. (b) Cells were cultured for 48 h in the presence of G, S, or C (either alone or in different combinations) and cell viability was monitored in an MTT assay. N = 6 of experiments performed at each time point. * P < 0.05 versus the control group; a P < 0.05 versus G; b P < 0.05 versus S; c P < 0.05 versus C; d P < 0.05 versus GS; e P < 0.05 versus GC; f P < 0.05 versus the SC; and # P < 0.05 versus the vehicle-treated control at the same time-point.

Article Snippet: The cardiomyoblast cell line, H9c2, was purchased from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Marker, Cell Culture, MTT Assay, Control

Journal: BioMed Research International

Article Title: Interleukin-27 Protects Cardiomyocyte-Like H9c2 Cells against Metabolic Syndrome: Role of STAT3 Signaling

doi: 10.1155/2015/689614

Figure Lengend Snippet: Effects of IL-27 on STAT3 activity and cytochrome c release from H9c2 cells. (a) IL-27 increased the activity of STAT3 in a time-dependent manner compared with that in control (vehicle-treated) cells. (b) Lactate dehydrogenase (LDH) release by control cells indicates spontaneous turnover during cell growth. Note that IL-27 had no effect on LDH release. (c) Changes in STAT3 activity after 48 h of treatment with GSC, IL-27, or GSC + IL-27. Note that IL-27 partially reversed the glucose, sodium, and cholesterol (GSC) induced reduction in STAT3 activity. (d) Effects of GSC and IL-27 on LDH release. Note that the GSC-mediated increase in LDH release was attenuated by IL-27. (e) Apoptosis was evaluated by assessing the cytosolic levels of cytochrome c (Cyto c). Western blots showing the expression of cytochrome c in three representative cultures. Equal amounts of protein (20 μ g per lane) were loaded. The bar graph shows changes in cytochrome c expression (as measured by densitometry) after normalization against actin. N = 6 of experiments performed at each time-point or in each group. AU, arbitrary units. * P < 0.05 versus the zero time-point or the control (C) group; # P < 0.05 versus the IL-27 group; + P < 0.05 versus the GSC group.

Article Snippet: The cardiomyoblast cell line, H9c2, was purchased from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Activity Assay, Control, Western Blot, Expressing

Journal: International Journal of Molecular and Cellular Medicine

Article Title: Melatonin Ameliorates 5-Fluorouracil-Induced Cytotoxicity and Apoptosis in H9c2 Cell Line: Insights into Cytoprotection and Anti-Apoptotic Mechanisms

doi: 10.22088/IJMCM.BUMS.14.3.843

Figure Lengend Snippet: 5-Fluorouracil (5-FU) induces apoptosis in H9c2 cardiomyoblasts primarily by activating the executioner caspases 3/7 and associated pro-apoptotic signaling pathways. In contrast, co-treatment with melatonin (MLT) counteracts this effect by modulating key survival signaling pathways. MLT activates the PI3K/Akt and Nrf2/ARE axes, which leads to the suppression of caspase 3/7 activity and a reduction in oxidative stress, ultimately resulting in decreased apoptosis and enhanced cell protection.

Article Snippet: The rat myocardial cell lines, H9c2, were purchased from the Pasteur Institute of Iran (Tehran, Iran).

Techniques: Protein-Protein interactions, Activity Assay

Journal: International Journal of Molecular and Cellular Medicine

Article Title: Melatonin Ameliorates 5-Fluorouracil-Induced Cytotoxicity and Apoptosis in H9c2 Cell Line: Insights into Cytoprotection and Anti-Apoptotic Mechanisms

doi: 10.22088/IJMCM.BUMS.14.3.843

Figure Lengend Snippet: Cytotoxicity of 5-FU in H9c2 cells. Cells were exposed to different concentrations of 5-Fluorouracil (5-FU) ranging from 0 µM to 400 µM for an incubation period of 48 h. At the end of the exposure, MTT cell viability was determined as explained in the materials and methods (A). The MTT assay results obtained from H9c2 treated cells with different concentrations of Melatonin (MLT) ranging from 0 µM to 400 µM for an incubation period of 48 h. The data represented are the mean ± SD of three identical experiments made in three replicates. The dashed lines represent a 50% decrease in the MTT assay results (B).

Article Snippet: The rat myocardial cell lines, H9c2, were purchased from the Pasteur Institute of Iran (Tehran, Iran).

Techniques: Incubation, MTT Assay

Journal: International Journal of Molecular and Cellular Medicine

Article Title: Melatonin Ameliorates 5-Fluorouracil-Induced Cytotoxicity and Apoptosis in H9c2 Cell Line: Insights into Cytoprotection and Anti-Apoptotic Mechanisms

doi: 10.22088/IJMCM.BUMS.14.3.843

Figure Lengend Snippet: The cell viability of H9c2 cell line treated with different concentrations of MLT and 5-FU for 48 h. The data are presented as mean ± SD of three independent each performed in three replicates. **p<0.03, ****p<0.0001 compared to cells treated with 5-FU 300 µM. ^^^p<0.001, ^^^^p<0.0001 compared to cells treated with 5-FU 400 µM. ####p<0.0001 compared among the treated groups with the highest viability percentages.

Article Snippet: The rat myocardial cell lines, H9c2, were purchased from the Pasteur Institute of Iran (Tehran, Iran).

Techniques:

Journal: International Journal of Molecular and Cellular Medicine

Article Title: Melatonin Ameliorates 5-Fluorouracil-Induced Cytotoxicity and Apoptosis in H9c2 Cell Line: Insights into Cytoprotection and Anti-Apoptotic Mechanisms

doi: 10.22088/IJMCM.BUMS.14.3.843

Figure Lengend Snippet: Flow cytometric analysis of 5-FU and/or MLT exposure using AnnexinV/PI staining. H9c2 cells were treated with different concentrations of test compounds. (a) control group cells, (b) Cells treated with 5-FU 400 µM, (c) cells co-treated with 5-FU 400 µM and MLT 50 µM, (d) cells co-treated with 5-FU 400 µM and MLT 100 µM (A). The apoptotic cells were measured by flow cytometry, and the percentage of apoptosis of cells labeled as Anx V (+) PI (−) was determined. (Q1) early apoptotic cells, (Q2) late apoptotic cells, (Q3) necrotic cells, and (Q4) viable cells (B).

Article Snippet: The rat myocardial cell lines, H9c2, were purchased from the Pasteur Institute of Iran (Tehran, Iran).

Techniques: Staining, Control, Flow Cytometry, Labeling

Journal: International Journal of Molecular and Cellular Medicine

Article Title: Melatonin Ameliorates 5-Fluorouracil-Induced Cytotoxicity and Apoptosis in H9c2 Cell Line: Insights into Cytoprotection and Anti-Apoptotic Mechanisms

doi: 10.22088/IJMCM.BUMS.14.3.843

Figure Lengend Snippet: Effect of 5-FU and MLT on the caspase-3/7 activity in H9c2 cardiomyocyte cell line in vitro . Caspase-3/7 activity was examined by using fluorescent assay kits. The values are in mean ± SD. *p<0.05 compared to cells treated with 5-FU 300 µM, ^p<0.05 and ^^^p<0.001, compared to cells treated with 5-FU 400 µM, #p<0.05 compared among the treated groups with the lowest caspase 3/7 activities.

Article Snippet: The rat myocardial cell lines, H9c2, were purchased from the Pasteur Institute of Iran (Tehran, Iran).

Techniques: Activity Assay, In Vitro, Fluorescence

Journal: International Journal of Molecular Medicine

Article Title: S-allyl-cysteine sulfoxide (alliin) alleviates myocardial infarction by modulating cardiomyocyte necroptosis and autophagy

doi: 10.3892/ijmm.2019.4351

Figure Lengend Snippet: Alliin protects cardiomyocytes against apoptosis and necroptosis. (A) Effect of alliin on the rate of hypoxia-induced apoptosis and necroptosis in H9c2 cells, assessed by flow cytometry. (B) Time course of cell death (Annexin V + cells). (C) Expression and (D) relative levels of cleaved caspase-3 RIP1, RIP3 and TRAF2 in hypoxia-induced H9c2 cells treated without or with alliin. The data presented in each panel are representative of at least three independent experiments and are presented as the mean ± SEM. * P<0.05, ** P<0.01, *** P<0.001. Ctl, control; PI, propidium iodide; RIP, receptor-interacting protein; TRAF2, tumor necrosis factor receptor-associated factor 2.

Article Snippet: H9c2 cells were obtained from ScienCell Research Laboratories and were cultured in ECM containing 5% FBS, and 1% penicillin/streptomycin (all Gibco; Thermo Fisher Scientific, Inc.) at 37°C in 5% CO 2 .

Techniques: Flow Cytometry, Expressing, Control

Journal: International Journal of Molecular Medicine

Article Title: S-allyl-cysteine sulfoxide (alliin) alleviates myocardial infarction by modulating cardiomyocyte necroptosis and autophagy

doi: 10.3892/ijmm.2019.4351

Figure Lengend Snippet: RNA sequencing analysis reveals a role for alliin in autophagy. (A) KEGG pathway analysis of proteins regulated by alliin during hypoxia. (B) Autophagy-related genes regulated by alliin identified by RNA sequencing analysis. (C) Levels of ATG4A, ATG4B, ATG4C, ATG4D, ATG9A and ATG16L2 in primary cardiomyocytes were detected by reverse transcription-quantitative PCR analysis. (D) Expression levels of Beclin 1, LC3, ATG7, p62 and PPARγ in H9c2 cells under hypoxia were analyzed by western blotting, and the (E) effect of alliin was reversed by autophagy inhibitor 3-MA. The data presented in each panel are representative of at least three independent experiments and are presented as the mean ± SEM. * P<0.05, ** P<0.01 and *** P<0.001. Ctl, control; LC3, microtubule-associated protein 1 light chain 3; ATG, autophagy-related; PPARγ, peroxisome proliferator-activated receptor γ.

Article Snippet: H9c2 cells were obtained from ScienCell Research Laboratories and were cultured in ECM containing 5% FBS, and 1% penicillin/streptomycin (all Gibco; Thermo Fisher Scientific, Inc.) at 37°C in 5% CO 2 .

Techniques: RNA Sequencing, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Control

Journal: Biochemistry and Biophysics Reports

Article Title: Diallyl disulfide ameliorates isoproterenol induced cardiac hypertrophy activating mitochondrial biogenesis via eNOS-Nrf2-Tfam pathway in rats

doi: 10.1016/j.bbrep.2015.11.008

Figure Lengend Snippet: Effect of garlic, DADS and NaHS on cellular nitric oxide level and mitochondrial biogenesis against isoproterenol treated H9C2 cells and its inhibition by L-NAME. (A) Nitric oxide level. (B) CO1 mRNA expression. (C) Cyb mRNA expression. Data were shown as mean±SEM ( N =3), * p <0.05 versus Con, # p <0.05 versus Iso.

Article Snippet: H9C2 cells were purchased from Lonza, USA and cultured using DMEM media containing10% fetal bovine serum (FBS).

Techniques: Inhibition, Expressing