Journal: Frontiers in Pharmacology

Article Title: Ivabradine Ameliorates Cardiac Diastolic Dysfunction in Diabetic Mice Independent of Heart Rate Reduction

doi: 10.3389/fphar.2021.696635

Figure Lengend Snippet: JNK and p38 MAPK mediate high glucose (HG)-induced fibrogenic protein upregulation in vitro (A) NRCFs were confirmed by vimentin and DAPI staining (original magnification × 400). Scale bars, 50 μm. Data were obtained from three independent experiments (B and C) NRCFs transfected with or without JNK or p38 MAPK siRNA were evaluated for the protein expression of JNK and p38 MAPK by western blotting (D) NRCFs were exposed to HG for 0–48 h and JNK/p38 MAPK phosphorylation and total JNK/p38 MAPK expression were determined by western blotting (E) NRCFs were transfected with or without JNK/p38 MAPK siRNA and then exposed to HG for the indicated periods of time. The protein expression of PCNA, α-SMA, TIMP2, collagen I, collagen III, and MMP2 was determined by western blotting. Data in (B–E) are presented as mean ± SEM ( n = 3). p -values in (B–D) were calculated using paired Student’s t -test. p -value in (E) was calculated using one-way ANOVA with Tukey multiple comparison test. * p < 0.05, compared to the normal group; # p < 0.05, compared to the HG group.

Article Snippet: Antibodies against collagen I and collagen III were obtained from the Biorbyt Corporation (Orwell Furlong, Cambridge, United Kingdom).

Techniques: In Vitro, Staining, Transfection, Expressing, Western Blot, Phospho-proteomics, Comparison

Journal: Frontiers in Pharmacology

Article Title: Ivabradine Ameliorates Cardiac Diastolic Dysfunction in Diabetic Mice Independent of Heart Rate Reduction

doi: 10.3389/fphar.2021.696635

Figure Lengend Snippet: JNK and p38 MAPK mediate the diabetes-induced increase in fibrogenic protein expression in vivo (A) Left ventricular fibroblasts isolated from mice were confirmed by vimentin staining. Data was obtained from three independent experiments (B and C) Isolated left ventricular fibroblasts from wild-type mice that were or were not injected with lentivirus via a tail vein were evaluated for JNK and p38 MAPK protein expression after 4 weeks (D) Left ventricular fibroblasts were isolated from mice and JNK and p38 MAPK phosphorylation and total JNK and p38 MAPK expression were determined by western blotting (E) Isolated left ventricular fibroblasts from diabetic mice that had or had not been injected with lentivirus via a tail vein were evaluated for PCNA, α-SMA, TIMP2, collagen I, collagen III, and MMP2 expression after 4 weeks (F) Between 7 and 11 weeks following the tail vein injection of lentivirus, the blood glucose concentration and body mass of the mice were measured. Data in (B–F) are presented as mean ± SEM ( n = 6). p -values in (B–D) were calculated using paired Student’s t -test. p -values in (E-F) were calculated using one-way ANOVA with Tukey multiple comparison test. * p < 0.05, compared to the Control group; # p < 0.05, compared to the db/db mice group.

Article Snippet: Antibodies against collagen I and collagen III were obtained from the Biorbyt Corporation (Orwell Furlong, Cambridge, United Kingdom).

Techniques: Expressing, In Vivo, Isolation, Staining, Injection, Phospho-proteomics, Western Blot, Concentration Assay, Comparison, Control

Journal: Frontiers in Pharmacology

Article Title: Ivabradine Ameliorates Cardiac Diastolic Dysfunction in Diabetic Mice Independent of Heart Rate Reduction

doi: 10.3389/fphar.2021.696635

Figure Lengend Snippet: Ivabradine, but not zatebradine, reduces JNK/p38 MAPK activation and the high glucose (HG)-induced increase in fibrogenic protein expression in vitro (A) NRCFs were pretreated with or without ivabradine at the indicated concentrations for 30 min, and then exposed to HG for 48 h. JNK and p38 MAPK phosphorylation and total JNK and p38 MAPK expression were determined by western blotting (B) NRCFs were treated as described in (A) and the protein expression of PCNA, α-SMA, TIMP2, collagen I, collagen III, and MMP2 was determined by western blotting (C and D) NRCFs were pretreated with or without zatebradine at the indicated concentrations for 30 min, and then exposed to HG for 48 h. The proteins described in (A) and (B) were quantified by western blotting (E and F) NRCFs were treated as described in (A) and (C) and the cell proliferation rate was determined using an MTT assay. Data are presented as mean ± SEM ( n = 3). p -values were calculated using one-way ANOVA with Tukey multiple comparison test. * p < 0.05, compared to the normal group; # p < 0.05, compared to the HG group.

Article Snippet: Antibodies against collagen I and collagen III were obtained from the Biorbyt Corporation (Orwell Furlong, Cambridge, United Kingdom).

Techniques: Activation Assay, Expressing, In Vitro, Phospho-proteomics, Western Blot, MTT Assay, Comparison

Journal: Frontiers in Pharmacology

Article Title: Ivabradine Ameliorates Cardiac Diastolic Dysfunction in Diabetic Mice Independent of Heart Rate Reduction

doi: 10.3389/fphar.2021.696635

Figure Lengend Snippet: Ivabradine, but not zatebradine, ameliorates the diabetes-induced increases in fibrogenic protein expression and fibrosis (A) Collagen I, collagen III, TIMP2, and MMP2 protein expression was determined by western blotting in left ventricular fibroblasts isolated from mice administered ivabradine at the indicated doses (B) Left ventricular fibroblasts were isolated from mice administered zatebradine at the indicated doses and the expression of the proteins listed in (A) was measured by western blotting (C and D) Mice were treated as described in (A) and (B) and the degree of cardiac fibrosis was determined by Massonʼs trichrome staining and immunohistochemistry. The columns show the differences in collagen accumulation (original magnification ×400). Scale bars, 50 μm. Data in (A–B) ( n = 3) and (C–D) ( n = 6) are presented as mean ± SEM. p -values were calculated using one-way ANOVA with Tukey multiple comparison test. * p < 0.05, compared to the Control group; # p < 0.05, compared to the db/db mouse group.

Article Snippet: Antibodies against collagen I and collagen III were obtained from the Biorbyt Corporation (Orwell Furlong, Cambridge, United Kingdom).

Techniques: Expressing, Western Blot, Isolation, Staining, Immunohistochemistry, Comparison, Control

Journal: Current Protocols

Article Title: Cryopreservation of Human Adult Ventricular Tissue for the Preparation of Viable Myocardial Slices

doi: 10.1002/cpz1.70068

Figure Lengend Snippet: Primary Antibodies

Article Snippet: Type III Collagen , 1:2000 , Southern Biotech , 1330‐01.

Techniques:

Journal: Current Protocols

Article Title: Cryopreservation of Human Adult Ventricular Tissue for the Preparation of Viable Myocardial Slices

doi: 10.1002/cpz1.70068

Figure Lengend Snippet: Extracellular matrix. Representative images of immunohistochemical analysis of fresh and cryopreserved cardiac tissue. Type I Collagen (Collagen‐I) is shown in orange (A) and Type III Collagen (Collagen‐III) is shown in yellow (B) . Counterstaining of nuclei was performed with DAPI (blue).

Article Snippet: Type III Collagen , 1:2000 , Southern Biotech , 1330‐01.

Techniques: Immunohistochemical staining