human cardiomyocytes ac16 cells  (Procell Inc)

Journal: Frontiers in Immunology

Article Title: Limonin alleviates pyroptosis and inflammatory responses in cardiomyocytes of myocardial ischemia-reperfusion mice by inhibiting the caspase-3/GSDME pathway

doi: 10.3389/fimmu.2026.1704424

Figure Lengend Snippet: Limonin inhibits pyroptosis in cardiomyocyte AC16 cells. (A) CCK8 was used to detect the effects of limonin on the viability of AC16 cells after incubation with different concentrations of limonin for 24 h, and the OGD/R cell model was established after incubation (n = 5). (B) CCK8 was used to detect the effects of limonin (50 μmol/L) and triclabendazole (40 μmol/L) treatment for 24 h followed by OGD/R modeling on AC16 cells (n = 5). (C) PI/Hoechst 33258 double staining was used to detect the changes in pyroptosis (×100, Scale bar: 50 μm, n = 5). (D) Bar chart for statistical analysis of PI/Hoechst 33258. (E) Flow cytometry was used to detect the changes in pyroptosis (n = 5). ns represents no statistical significance compared with the control group, and * represents comparison between two groups (* P < 0.05, ** P < 0.01, *** P < 0.001*, **** P < 0.0001).

Article Snippet: Human cardiomyocytes AC16 cells were purchased from Wuhan Procell Life Science & Technology Co. Ltd. (Wuhan, China).

Techniques: Incubation, Double Staining, Flow Cytometry, Control, Comparison

Journal: Frontiers in Immunology

Article Title: Limonin alleviates pyroptosis and inflammatory responses in cardiomyocytes of myocardial ischemia-reperfusion mice by inhibiting the caspase-3/GSDME pathway

doi: 10.3389/fimmu.2026.1704424

Figure Lengend Snippet: Limonin can improve tissue damage and inflammatory response in I/R mice. (A) TUNEL staining was used to detect the changes in myocardial tissue apoptosis (×100, Scale bar: 100 μm, n = 10). (B) HE staining was used to detect pathological changes in the myocardial tissue (×100, Scale bar: 100 μm, n = 10). (C) Masson staining was used to observe changes in the myocardial tissue (×100, Scale bar: 100 μm, n = 10). (D) ELISA was used to detect changes in inflammatory factors IL-6, IL-1β, TNF-α, and IL-10 in the supernatant of AC16 cells (n = 5). (E) ELISA was also used to detect changes in inflammatory factors IL-6, LI-1β, TNF-α, and IL-10 in mouse blood (n = 10). (F) Relative expression levels of IL-6, IL-1β, TNF-α, and IL-10 mRNA in mouse myocardial tissues detected by RT-qPCR. (G) Relative expression levels of IL-6, IL-1β, TNF-α, and IL-10 mRNA in AC16 cells detected by RT-qPCR. * represents comparison between two groups (* P < 0.05, ** P < 0.01, *** P < 0.001*, **** P < 0.0001).

Article Snippet: Human cardiomyocytes AC16 cells were purchased from Wuhan Procell Life Science & Technology Co. Ltd. (Wuhan, China).

Techniques: TUNEL Assay, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Comparison

Journal: Frontiers in Immunology

Article Title: Limonin alleviates pyroptosis and inflammatory responses in cardiomyocytes of myocardial ischemia-reperfusion mice by inhibiting the caspase-3/GSDME pathway

doi: 10.3389/fimmu.2026.1704424

Figure Lengend Snippet: Limonin can inhibit the Caspase-3/GSDME signaling pathway. (A) Relative expression levels of caspase-3, cleaved-caspase-3, GSDME-F, GSDME-N, and IL-18 proteins in myocardial tissues detected by western blot experiment (n = 5). (B) Relative expression levels of caspase-3, cleaved-caspase-3, GSDME-F, GSDME-N, and IL-18 proteins in AC16 cells detected by western blot experiment (n = 5). (C) Detection of GSDME-N expression levels in AC16 cells by immunofluorescence assay (200×, scale bar: 50 μm, n = 5) (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: Human cardiomyocytes AC16 cells were purchased from Wuhan Procell Life Science & Technology Co. Ltd. (Wuhan, China).

Techniques: Expressing, Western Blot, Immunofluorescence

Journal: Cardiovascular Toxicology

Article Title: Taurine Alleviates Sepsis-Induced Myocardial Injury by Inhibiting NF-κB Pathway-Mediated Inflammation, Mitochondrial Dysfunction, and Apoptosis

doi: 10.1007/s12012-026-10096-w

Figure Lengend Snippet: Taurine reduces inflammation and oxidative stress in CLP-induced SICM mice. A – C Serum levels of IL-1β, TNF-α, and IL-6 at 24 h post-surgery ( n = 6). D – F Myocardial tissue mRNA levels of IL-1β, TNF-α, and IL-6 at 24 h post-surgery ( n = 6). G – I Myocardial tissue levels of MDA, SOD, and GSH-Px at 24 h post-surgery ( n = 3). J – K Representative Western blot images and quantitative analysis of Nrf2. L AC16 cells viability treated with LPS (10 µg/ml) and different concentrations of taurine (0, 10, 20, 40, 80 mM) for 24 h ( n = 3). M , N Representative fluorescence images and quantification of relative fluorescence intensity of intracellular ROS ( n = 3). Data shown are Mean ± SEM

Article Snippet: The AC16 Human Cardiomyocyte cells (Servicebio, Wuhan, China) were cultured in high-glucose Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) at 37 °C in a humidified atmosphere containing 5% CO2.

Techniques: Western Blot, Fluorescence

Journal: Cardiovascular Toxicology

Article Title: Taurine Alleviates Sepsis-Induced Myocardial Injury by Inhibiting NF-κB Pathway-Mediated Inflammation, Mitochondrial Dysfunction, and Apoptosis

doi: 10.1007/s12012-026-10096-w

Figure Lengend Snippet: Effects of taurine on myocardial mitochondrial ultrastructure and mitochondrial function. A , B Representative transmission electron microscopy images of myocardial mitochondria and Flameng scores. Each symbol represents the average mitochondrial score per field of view (FOV). Each group has three mice and five FOVs were randomly selected for assay from each anima (scale bar = 2 μm/0.5 μm). C OCR curve of AC16 cells (normalized to 10,000 cells). (D-F) Quantitative analysis of basal respiration D , maximal respiration E and ATP yield F calculated from OCR data, expressed in pmol/min/10,000 cells ( n = 3). G Representative images of JC-1 staining of cells used to measure MMP after different treatments. H Quantification of cellular MMP levels using ImageJ software ( n = 3). Data shown are Mean ± SEM

Article Snippet: The AC16 Human Cardiomyocyte cells (Servicebio, Wuhan, China) were cultured in high-glucose Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) at 37 °C in a humidified atmosphere containing 5% CO2.

Techniques: Transmission Assay, Electron Microscopy, Staining, Software

Journal: Cardiovascular Toxicology

Article Title: Taurine Alleviates Sepsis-Induced Myocardial Injury by Inhibiting NF-κB Pathway-Mediated Inflammation, Mitochondrial Dysfunction, and Apoptosis

doi: 10.1007/s12012-026-10096-w

Figure Lengend Snippet: Effects of taurine on apoptosis in myocardial tissue and cardiomyocytes. A , B Representative images of TUNEL staining (apoptotic cells were showing green) in heart tissues sections and quantitative analysis (scale bar = 50 μm; n = 3). C , D Representative flow cytometry plots showing apoptosis levels and its quantification in AC16 cells ( n = 3). E – H apoptosis-related proteins (Bcl-2, Bax, Cleaved-Caspase-3) in heart tissues ( n = 3). Data shown are Mean ± SEM

Article Snippet: The AC16 Human Cardiomyocyte cells (Servicebio, Wuhan, China) were cultured in high-glucose Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) at 37 °C in a humidified atmosphere containing 5% CO2.

Techniques: TUNEL Assay, Staining, Flow Cytometry

Journal: Cardiovascular Toxicology

Article Title: Taurine Alleviates Sepsis-Induced Myocardial Injury by Inhibiting NF-κB Pathway-Mediated Inflammation, Mitochondrial Dysfunction, and Apoptosis

doi: 10.1007/s12012-026-10096-w

Figure Lengend Snippet: Taurine inhibits SICM-induced activation of the NF-κB pathway. A Representative immunofluorescence images showing NF-κB p65 (green;scale bar = 50 μm) nuclear translocation in AC16 cells after LPS stimulation (10 µg/ml, 24 h). Nuclei are stained with DAPI (blue;scale bar = 50 μm). B , C Representative Western blot images and quantitative analysis of p-NF-κB p65, NF-κB p65, p-IKBα and IKBα protein levels in mice cardiac muscles ( n = 3). Data shown are Mean ± SEM

Article Snippet: The AC16 Human Cardiomyocyte cells (Servicebio, Wuhan, China) were cultured in high-glucose Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) at 37 °C in a humidified atmosphere containing 5% CO2.

Techniques: Activation Assay, Immunofluorescence, Translocation Assay, Staining, Western Blot, Muscles

Journal: Cardiovascular Toxicology

Article Title: Taurine Alleviates Sepsis-Induced Myocardial Injury by Inhibiting NF-κB Pathway-Mediated Inflammation, Mitochondrial Dysfunction, and Apoptosis

doi: 10.1007/s12012-026-10096-w

Figure Lengend Snippet: NF-κB activation weakened the protective effect of taurine on LPS-induced AC16 cells. A , B Representative fluorescence images and quantification of relative fluorescence intensity of intracellular ROS ( n = 3). C , D Representative images and quantification of JC-1 staining of cells used to measure MMP after different treatments ( n = 3). E , F Representative flow cytometry plots showing apoptosis levels and its quantification in AC16 cells ( n = 3). G – J Representative Western blot images and quantification of apoptosis-related proteins (Bcl-2, Bax, Cleaved-Caspase-3) in heart tissues ( n = 3). Data shown are Mean ± SEM

Article Snippet: The AC16 Human Cardiomyocyte cells (Servicebio, Wuhan, China) were cultured in high-glucose Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) at 37 °C in a humidified atmosphere containing 5% CO2.

Techniques: Activation Assay, Fluorescence, Staining, Flow Cytometry, Western Blot

Journal: Arquivos Brasileiros de Cardiologia

Article Title: Knockdown do RNA Longo Não Codificante ZEB1-AS1 Acelera a Hipertrofia Cardíaca pela Via miR-186-5p/HDAC2

doi: 10.36660/abc.20240703

Figure Lengend Snippet: – Superexpressão de ZEB1-AS1 em tecidos com HVE e em células AC16 estimuladas com ISO. A) Expressão de ZEB1-AS1 em corações com HVE (n = 20) versus doadores normais (n = 10) por qRT-PCR; B) Área de superfície de células AC16 após tratamento com PBS ou ISO, avaliada por coloração por imunofluorescência; C) Níveis de mRNA de BNP, ANP e β-MHC em células AC16 após PBS ou ISO por qRT-PCR; D) Níveis de proteína de BNP, ANP e β-MHC em células AC16 após PBS ou ISO por Western blot; E) Expressão de ZEB1-AS1 em células AC16 após PBS ou ISO por qRT-PCR. Todos os experimentos foram realizados em triplicata. *p < 0,05, *p < 0,01. ANP: peptídeo natriurético atrial; BNP: peptídeo natriurético tipo B; β-MHC: cadeia pesada de miosina β; HC: hipertrofia cardíaca; HVE: hipertrofia ventricular esquerda; ISO: isoproterenol; PBS: solução salina tamponada com fosfato; qRT-PCR: reação em cadeia da polimerase quantitativa em tempo real.

Article Snippet: Human cardiomyocyte cell line AC16 was obtained from the American Type Culture Collection (USA).

Techniques: Quantitative RT-PCR, Western Blot

Journal: Arquivos Brasileiros de Cardiologia

Article Title: Knockdown do RNA Longo Não Codificante ZEB1-AS1 Acelera a Hipertrofia Cardíaca pela Via miR-186-5p/HDAC2

doi: 10.36660/abc.20240703

Figure Lengend Snippet: – A deficiência de ZEB1-AS1 atenua a HC induzida por ISO em células AC16. A) Expressão de ZEB1-AS1 em células AC16 transfectadas com sh-NC ou sh-ZEB1-AS1 por qRT-PCR; B) área de superfície das AC16 após estimulação com ISO com sh-NC ou sh-ZEB1-AS1, avaliada por coloração por imunofluorescência; C) níveis de mRNA de BNP, ANP e β-MHC em AC16 estimuladas com ISO com sh-NC ou sh-ZEB1-AS1 por qRT-PCR; D) níveis de proteína de BNP, ANP e β-MHC em AC16 estimuladas com ISO com sh-NC ou sh-ZEB1-AS1 por Western blot. Todos os experimentos foram realizados em triplicata. *p < 0,05, *p < 0,01. ANP: peptídeo natriurético atrial; BNP: peptídeo natriurético tipo B; β-MHC: cadeia pesada de miosina β; HC: hipertrofia cardíaca; ISO: isoproterenol; qRT-PCR: reação em cadeia da polimerase quantitativa em tempo real; sh-NC: short hairpin RNA–negative control; sh-ZEB1-AS1: shRNA direcionado a ZEB1-AS1.

Article Snippet: Human cardiomyocyte cell line AC16 was obtained from the American Type Culture Collection (USA).

Techniques: Quantitative RT-PCR, Western Blot, shRNA, Negative Control

Journal: Arquivos Brasileiros de Cardiologia

Article Title: Knockdown do RNA Longo Não Codificante ZEB1-AS1 Acelera a Hipertrofia Cardíaca pela Via miR-186-5p/HDAC2

doi: 10.36660/abc.20240703

Figure Lengend Snippet: – ZEB1-AS1 promove a HC ao regular a via miR-186-5p/HDAC2. A) mRNA de HDAC2 em células AC16 estimuladas com ISO e transfectadas com os construtos indicados, medido por qRT-PCR; B) área de superfície das AC16 após estimulação com ISO e transfecção com os construtos indicados, avaliada por coloração por imunofluorescência; C) níveis de mRNA de BNP, ANP e β-MHC em AC16 estimuladas com ISO após transfecção, medidos por qRT-PCR; D) níveis de proteína de BNP, ANP e β-MHC em AC16 estimuladas com ISO após transfecção, medidos por Western blot. Todos os experimentos foram realizados em triplicata. *p < 0,05, *p < 0,01. ANP: peptídeo natriurético atrial; BNP: peptídeo natriurético tipo B; β-MHC: cadeia pesada de miosina β; HC: hipertrofia cardíaca; HDAC2: histone deacetylase 2; ISO: isoproterenol; qRT-PCR: reação em cadeia da polimerase quantitativa em tempo real; sh-NC: short hairpin RNA–negative control; sh-ZEB1-AS1: zinc finger E-box binding homeobox 1 antisense RNA 1 direcionado ao short hairpin RNA.

Article Snippet: Human cardiomyocyte cell line AC16 was obtained from the American Type Culture Collection (USA).

Techniques: Quantitative RT-PCR, Western Blot, Histone Deacetylase Assay, shRNA, Negative Control, Binding Assay

Journal: Arquivos Brasileiros de Cardiologia

Article Title: Knockdown do RNA Longo Não Codificante ZEB1-AS1 Acelera a Hipertrofia Cardíaca pela Via miR-186-5p/HDAC2

doi: 10.36660/abc.20240703

Figure Lengend Snippet: – Upregulation of ZEB1-AS1 in LVH tissues and ISO-stimulated AC16 cells. A) ZEB1-AS1 expression in LVH hearts (n = 20) versus normal donors (n = 10) by qRT-PCR. B) AC16 cell surface area after PBS or ISO treatment assessed by immunofluorescence. C) mRNA levels of BNP, ANP, and β-MHC in AC16 cells after PBS or ISO by qRT-PCR. D) Protein levels of BNP, ANP, and β-MHC in AC16 cells after PBS or ISO by Western blot. E) ZEB1-AS1 expression in AC16 cells after PBS or ISO by qRT-PCR. All experiment was performed in triplicate. *p < 0.05, **p < 0.01. ANP: atrial natriuretic peptide; BNP: B-type natriuretic peptide; β-MHC: beta-myosin heavy chain; CH: cardiac hypertrophy; ISO: isoproterenol; LVH: left ventricular hypertrophy; PBS: phosphate-buffered saline; qRT-PCR: quantitative real-time polymerase chain reaction.

Article Snippet: Human cardiomyocyte cell line AC16 was obtained from the American Type Culture Collection (USA).

Techniques: Expressing, Quantitative RT-PCR, Immunofluorescence, Western Blot, Saline, Real-time Polymerase Chain Reaction

Journal: Arquivos Brasileiros de Cardiologia

Article Title: Knockdown do RNA Longo Não Codificante ZEB1-AS1 Acelera a Hipertrofia Cardíaca pela Via miR-186-5p/HDAC2

doi: 10.36660/abc.20240703

Figure Lengend Snippet: – ZEB1-AS1 deficiency mitigates ISO-induced CH in AC16 cells. A) ZEB1-AS1 expression in AC16 cells transfected with sh-NC or sh-ZEB1-AS1 by qRT-PCR; B) AC16 cell surface area after ISO stimulation with sh-NC or sh-ZEB1-AS1 assessed by immunofluorescence; C) mRNA levels of BNP, ANP, and β-MHC in ISO-stimulated AC16 cells with sh-NC or sh-ZEB1-AS1 by qRT-PCR; D) protein levels of BNP, ANP, and β-MHC in ISO-stimulated AC16 cells with sh-NC or sh-ZEB1-AS1 by Western blot. All experiments were performed in triplicate. *p < 0.05, **p < 0.01. ANP: atrial natriuretic peptide; BNP: B-type natriuretic peptide; β-MHC: beta-myosin heavy chain; CH: cardiac hypertrophy; ISO: isoproterenol; qRT-PCR: quantitative real-time polymerase chain reaction; sh-NC: short hairpin RNA–negative control; sh-ZEB1-AS1: ZEB1-AS1-targeting shRNA.

Article Snippet: Human cardiomyocyte cell line AC16 was obtained from the American Type Culture Collection (USA).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Immunofluorescence, Western Blot, Real-time Polymerase Chain Reaction, shRNA, Negative Control

Journal: Arquivos Brasileiros de Cardiologia

Article Title: Knockdown do RNA Longo Não Codificante ZEB1-AS1 Acelera a Hipertrofia Cardíaca pela Via miR-186-5p/HDAC2

doi: 10.36660/abc.20240703

Figure Lengend Snippet: – ZEB1-AS1 promotes CH by regulating the miR-186-5p/HDAC2 pathway. A) HDAC2 mRNA in ISO-stimulated AC16 cells transfected with the indicated constructs measured by qRT-PCR; B) AC16 cell surface area after ISO stimulation and transfection with the indicated constructs assessed by immunofluorescence; C) mRNA levels of BNP, ANP, and β-MHC in ISO-stimulated AC16 cells after transfection measured by qRT-PCR; D) protein levels of BNP, ANP, and β-MHC in ISO-stimulated AC16 cells after transfection measured by Western blot. All experiments were performed in triplicate. *p < 0.05, **p < 0.01. ANP: atrial natriuretic peptide; BNP: B-type natriuretic peptide; β-MHC: beta-myosin heavy chain; CH: cardiac hypertrophy; HDAC2: histone deacetylase 2; ISO: isoproterenol; qRT-PCR: quantitative real-time polymerase chain reaction; sh-NC: short hairpin RNA–negative control; sh-ZEB1-AS1: zinc finger E-box binding homeobox 1 antisense RNA 1-targeting short hairpin RNA; miR-186-5p: microRNA-186-5p.

Article Snippet: Human cardiomyocyte cell line AC16 was obtained from the American Type Culture Collection (USA).

Techniques: Transfection, Construct, Quantitative RT-PCR, Immunofluorescence, Western Blot, Histone Deacetylase Assay, Real-time Polymerase Chain Reaction, shRNA, Negative Control, Binding Assay