Journal: Acta Physiologica (Oxford, England)

Article Title: Angiotensin‐(1–7) Alleviates Isoproterenol‐Induced Cardiac Hypertrophy by Suppressing Autophagy and Apoptosis Through the Synergistic Action of Mas Receptor and Angiotensin II Type 2 Receptor

doi: 10.1111/apha.70200

Figure Lengend Snippet: Physical and chemical properties and biosafety assessment of Ang‐(1–7). (A) Key molecular properties: molecular weight (MW), net charge, isoelectric point (pI), hydrophobicity (GRAVY index), and aromaticity. (B, C) Topological structure predicted by Deep TMHMM, showing a spherical soluble conformation (no transmembrane helices) and predicted functional sites (red‐intracellular; blue‐extracellular). (D) Hemolytic activity ( n = 6). (E) Cytotoxicity assessment in H9c2, HepG2, and NRK‐52E cell lines ( n = 6). Cell viability was determined using the Cell Counting Kit‐8 (CCK‐8) assay. The data is expressed as an mean ± standard deviation (SD).

Article Snippet: After 7 days' acclimatization, animals were randomized (blinded assessment) into five groups ( n = 6 per group) receiving daily Subcutaneous injection for 7 days [ , , ]:Ctrl: saline (Sanlian, Harbin, China); ISO: isoproterenol 5 mg/kg/day (Solarbio, II0200, Beijing, China; dissolved in DMSO); ISO + Ang‐(1–7): Ang‐(1–7) 576 μg/kg/day (MCE, HY‐12403, USA; in distilled water); ISO + Ang‐(1–7) + A‐779: A‐7791148 μg/kg/day (MCE, HY‐P0216, USA); ISO + Ang‐(1–7) + PD123319 : PD123319 5 mg/kg/day (MCE, HY‐10259A, USA); ISO + Ang‐(1–7) + PD123319 + A779.

Techniques: Molecular Weight, Functional Assay, Activity Assay, Cell Counting, CCK-8 Assay, Standard Deviation

Journal: Acta Physiologica (Oxford, England)

Article Title: Angiotensin‐(1–7) Alleviates Isoproterenol‐Induced Cardiac Hypertrophy by Suppressing Autophagy and Apoptosis Through the Synergistic Action of Mas Receptor and Angiotensin II Type 2 Receptor

doi: 10.1111/apha.70200

Figure Lengend Snippet: Effects of Ang‐(1–7) on isoproterenol (ISO)‐induced cardiac hypertrophy and fibrosis. (A) HE staining and Masson staining of myocardial sections (scale bar = 20 μm). (B, C) Quantitative analysis of myocardial cell cross‐sectional area and collagen volume fraction. (D, E) Macroscopic morphology and cross‐sectional observation of the heart, with measurement of heart weight/body weight ratio (HW/BW) (scale bar = 2 mm). (F) Western blot analysis of ANP, BNP, and β‐MHC expression. (G–I) Quantitative analysis of band intensity normalized to GAPDH. The data is expressed as an mean ± standard deviation (SD) ( n = 5). A + I, Ang‐(1–7) + ISO; AntM+A + I, A‐779 + Ang‐(1–7) + ISO; AntA+A + I, PD123319 + Ang‐(1–7) + ISO. *** p < 0.001 versus Ctrl; ### p < 0.001 versus ISO; &&& p < 0.001, && p < 0.01, & p < 0.05 versus Ang‐(1–7) + ISO.

Article Snippet: After 7 days' acclimatization, animals were randomized (blinded assessment) into five groups ( n = 6 per group) receiving daily Subcutaneous injection for 7 days [ , , ]:Ctrl: saline (Sanlian, Harbin, China); ISO: isoproterenol 5 mg/kg/day (Solarbio, II0200, Beijing, China; dissolved in DMSO); ISO + Ang‐(1–7): Ang‐(1–7) 576 μg/kg/day (MCE, HY‐12403, USA; in distilled water); ISO + Ang‐(1–7) + A‐779: A‐7791148 μg/kg/day (MCE, HY‐P0216, USA); ISO + Ang‐(1–7) + PD123319 : PD123319 5 mg/kg/day (MCE, HY‐10259A, USA); ISO + Ang‐(1–7) + PD123319 + A779.

Techniques: Staining, Western Blot, Expressing, Standard Deviation

Journal: Acta Physiologica (Oxford, England)

Article Title: Angiotensin‐(1–7) Alleviates Isoproterenol‐Induced Cardiac Hypertrophy by Suppressing Autophagy and Apoptosis Through the Synergistic Action of Mas Receptor and Angiotensin II Type 2 Receptor

doi: 10.1111/apha.70200

Figure Lengend Snippet: Ang‐(1–7) improves ISO‐induced ventricular remodeling and dysfunction via MasR and AT 2 R. (A) Representative M‐mode echocardiograms from each group (scale bar = 2 mm). (B, C) Quantification of left ventricular internal diameters at systole (LVIDs) and diastole (LVIDd). (D, E) Left ventricular ejection fraction (LVEF%) and fractional shortening (LVFS%). (F, G) Left ventricular posterior wall thickness at systole (LVPWs) and diastole (LVPWd). (H) Representative immunofluorescence images of cardiomyocytes (green, α‐actinin; blue, nuclei; scale bar = 10 μm). (I) Relative mRNA expression of ANP, BNP, and β‐MHC ( n = 6). The data is expressed as an mean ± standard deviation (SD) ( n = 5). A + I, Ang‐(1–7) + ISO; AntM + A + I, A‐779 + Ang‐(1–7) + ISO; AntA + A + I, PD123319 + Ang‐(1–7) + ISO; AntM + AntA + A + I, A‐779 + PD123319 + Ang‐(1–7) + ISO. *** p < 0.001, ** p < 0.01 versus Ctrl; ### p < 0.001, ## p < 0.01 versus ISO; &&& p < 0.001, && p < 0.01, & p < 0.05 versus Ang‐(1–7) + ISO, & p < 0.05 versus ISO.

Article Snippet: After 7 days' acclimatization, animals were randomized (blinded assessment) into five groups ( n = 6 per group) receiving daily Subcutaneous injection for 7 days [ , , ]:Ctrl: saline (Sanlian, Harbin, China); ISO: isoproterenol 5 mg/kg/day (Solarbio, II0200, Beijing, China; dissolved in DMSO); ISO + Ang‐(1–7): Ang‐(1–7) 576 μg/kg/day (MCE, HY‐12403, USA; in distilled water); ISO + Ang‐(1–7) + A‐779: A‐7791148 μg/kg/day (MCE, HY‐P0216, USA); ISO + Ang‐(1–7) + PD123319 : PD123319 5 mg/kg/day (MCE, HY‐10259A, USA); ISO + Ang‐(1–7) + PD123319 + A779.

Techniques: Immunofluorescence, Expressing, Standard Deviation

Journal: Acta Physiologica (Oxford, England)

Article Title: Angiotensin‐(1–7) Alleviates Isoproterenol‐Induced Cardiac Hypertrophy by Suppressing Autophagy and Apoptosis Through the Synergistic Action of Mas Receptor and Angiotensin II Type 2 Receptor

doi: 10.1111/apha.70200

Figure Lengend Snippet: Ang‐(1–7) regulates the expression of MasR and AT 2 R in H9c2 cardiomyocytes and exhibits cross‐inhibition with receptor antagonists. (A) Immunofluorescence images and immunofluorescence intensity of MasR (green), AT 2 R (red), and DAPI (blue) in H9c2 cells from different treatment groups, reflecting receptor expression levels (scale bar = 80 μm) ( n = 5). (B) Shows the expression of AT 2 R and MasR mRNA in H9c2 cells ( n = 6). (C) Analyzes MasR expression and relative thermal stability in cardiomyocytes at different temperatures (37°C–58°C). The data is expressed as an mean ± standard deviation (SD) ( n = 5). A + I, Ang‐(1–7) + ISO; AntM + A + I, A‐779 + Ang‐(1–7) + ISO; AntA + A + I, PD123319 + Ang‐(1–7) + ISO; AntM + AntA + A + I, A‐779 + PD123319 + Ang‐(1–7) + ISO. *** p < 0.001, * p < 0.05 versus Ctrl; #### p < 0.0001, ### p < 0.001 versus ISO; &&& p < 0.001, && p < 0.01, & p < 0.05 versus Ang‐(1–7) + ISO.

Article Snippet: After 7 days' acclimatization, animals were randomized (blinded assessment) into five groups ( n = 6 per group) receiving daily Subcutaneous injection for 7 days [ , , ]:Ctrl: saline (Sanlian, Harbin, China); ISO: isoproterenol 5 mg/kg/day (Solarbio, II0200, Beijing, China; dissolved in DMSO); ISO + Ang‐(1–7): Ang‐(1–7) 576 μg/kg/day (MCE, HY‐12403, USA; in distilled water); ISO + Ang‐(1–7) + A‐779: A‐7791148 μg/kg/day (MCE, HY‐P0216, USA); ISO + Ang‐(1–7) + PD123319 : PD123319 5 mg/kg/day (MCE, HY‐10259A, USA); ISO + Ang‐(1–7) + PD123319 + A779.

Techniques: Expressing, Inhibition, Immunofluorescence, Standard Deviation

Journal: Acta Physiologica (Oxford, England)

Article Title: Angiotensin‐(1–7) Alleviates Isoproterenol‐Induced Cardiac Hypertrophy by Suppressing Autophagy and Apoptosis Through the Synergistic Action of Mas Receptor and Angiotensin II Type 2 Receptor

doi: 10.1111/apha.70200

Figure Lengend Snippet: Ang‐(1–7) regulates the expression of MasR and AT 2 R and their heterodimer formation. (A) Immunofluorescence staining images of MasR (green), AT 2 R (red), and DAPI (blue) in myocardial tissue from mice in each treatment group. Arrows indicate the localization of receptors in the myocardium (scale bar = 20 μm) ( n = 5). (B) Protein expression and quantitative analysis of MasR and AT 2 R in myocardial tissue ( n = 6). (C) Molecular docking simulations showing the interactions between MasR and AT 2 R, as well as between Ang‐(1–7) and the receptors, along with the binding energies (ΔG) for each interaction. (D) Immunoprecipitation (Co‐IP): Interaction between MasR and AT 2 R in cardiac muscle tissue. The data is expressed as an mean ± standard deviation (SD). A + I, Ang‐(1–7) + ISO; AntM + A + I, A‐779 + Ang‐(1–7) + ISO; Ant A + A + I, PD123319 + Ang‐(1–7) + ISO. * p < 0.05 versus Ctrl; ### p < 0.001 versus ISO; &&& p < 0.001, & p < 0.05 versus Ang‐(1–7) + ISO.

Article Snippet: After 7 days' acclimatization, animals were randomized (blinded assessment) into five groups ( n = 6 per group) receiving daily Subcutaneous injection for 7 days [ , , ]:Ctrl: saline (Sanlian, Harbin, China); ISO: isoproterenol 5 mg/kg/day (Solarbio, II0200, Beijing, China; dissolved in DMSO); ISO + Ang‐(1–7): Ang‐(1–7) 576 μg/kg/day (MCE, HY‐12403, USA; in distilled water); ISO + Ang‐(1–7) + A‐779: A‐7791148 μg/kg/day (MCE, HY‐P0216, USA); ISO + Ang‐(1–7) + PD123319 : PD123319 5 mg/kg/day (MCE, HY‐10259A, USA); ISO + Ang‐(1–7) + PD123319 + A779.

Techniques: Expressing, Immunofluorescence, Staining, Binding Assay, Immunoprecipitation, Co-Immunoprecipitation Assay, Standard Deviation

Journal: Acta Physiologica (Oxford, England)

Article Title: Angiotensin‐(1–7) Alleviates Isoproterenol‐Induced Cardiac Hypertrophy by Suppressing Autophagy and Apoptosis Through the Synergistic Action of Mas Receptor and Angiotensin II Type 2 Receptor

doi: 10.1111/apha.70200

Figure Lengend Snippet: Ang‐(1–7) attenuates ISO‐induced excessive autophagy and apoptosis in vivo by regulating MasR and AT 2 R. (A) Transmission electron microscopy (TEM) images showing autophagosomes, lysosomes, and damaged mitochondria in cardiomyocytes; quantification of damaged mitochondria is shown (scale bar = 5 μm, 2 μm n = 4). (B) Western blot analysis of autophagy‐related proteins LC3‐II/I, Beclin1, and P62 ( n = 5). (C) Western blot analysis of apoptosis‐related proteins Bcl‐2, Bax, and cleaved caspase‐3 ( n = 5). (D, E) Quantitative densitometry analysis of autophagy‐ and apoptosis‐related proteins normalized to GAPDH. The data is expressed as an mean ± standard deviation (SD). A + I, Ang‐(1–7) + ISO; AntM + A + I, A‐779 + Ang‐(1–7) + ISO; Ant A + A + I, PD123319 + Ang‐(1–7) + ISO. *** p < 0.001 versus Ctrl; ### p < 0.001, # p < 0.05 versus ISO; &&& p < 0.001, && p < 0.01, & p < 0.05 versus Ang‐(1–7) + ISO.

Article Snippet: After 7 days' acclimatization, animals were randomized (blinded assessment) into five groups ( n = 6 per group) receiving daily Subcutaneous injection for 7 days [ , , ]:Ctrl: saline (Sanlian, Harbin, China); ISO: isoproterenol 5 mg/kg/day (Solarbio, II0200, Beijing, China; dissolved in DMSO); ISO + Ang‐(1–7): Ang‐(1–7) 576 μg/kg/day (MCE, HY‐12403, USA; in distilled water); ISO + Ang‐(1–7) + A‐779: A‐7791148 μg/kg/day (MCE, HY‐P0216, USA); ISO + Ang‐(1–7) + PD123319 : PD123319 5 mg/kg/day (MCE, HY‐10259A, USA); ISO + Ang‐(1–7) + PD123319 + A779.

Techniques: In Vivo, Transmission Assay, Electron Microscopy, Western Blot, Standard Deviation

Journal: Acta Physiologica (Oxford, England)

Article Title: Angiotensin‐(1–7) Alleviates Isoproterenol‐Induced Cardiac Hypertrophy by Suppressing Autophagy and Apoptosis Through the Synergistic Action of Mas Receptor and Angiotensin II Type 2 Receptor

doi: 10.1111/apha.70200

Figure Lengend Snippet: Ang‐(1–7) regulates autophagic flux and apoptosis in H9c2 cardiomyocytes through MasR and AT 2 R. (A) Representative images of TUNEL immunofluorescence staining (green fluorescence labels apoptotic cells) and Hoechst nuclear staining (blue) ( n = 6), with the Merge panel showing their overlay. (B) Western blot analysis of apoptosis related proteins (Bax, Bcl 2 , Cleaved caspase‐3) expression levels, with GAPDH as the internal control protein ( n = 3). (C) Western blot analysis of autophagy‐related proteins (LC3‐II, Beclin1, P62) expression levels, with GAPDH as the internal control protein ( n = 3). The data is expressed as an mean ± standard deviation (SD). A + I, Ang‐(1–7) + ISO; AntM + A + I, A‐779 + Ang‐(1–7) + ISO; Ant A + A + I, PD123319 + Ang‐(1–7) + ISO; AntM + AntA + A + I, A‐779 + PD123319 + Ang‐(1–7) + ISO.

Article Snippet: After 7 days' acclimatization, animals were randomized (blinded assessment) into five groups ( n = 6 per group) receiving daily Subcutaneous injection for 7 days [ , , ]:Ctrl: saline (Sanlian, Harbin, China); ISO: isoproterenol 5 mg/kg/day (Solarbio, II0200, Beijing, China; dissolved in DMSO); ISO + Ang‐(1–7): Ang‐(1–7) 576 μg/kg/day (MCE, HY‐12403, USA; in distilled water); ISO + Ang‐(1–7) + A‐779: A‐7791148 μg/kg/day (MCE, HY‐P0216, USA); ISO + Ang‐(1–7) + PD123319 : PD123319 5 mg/kg/day (MCE, HY‐10259A, USA); ISO + Ang‐(1–7) + PD123319 + A779.

Techniques: TUNEL Assay, Immunofluorescence, Staining, Fluorescence, Western Blot, Expressing, Control, Standard Deviation

Journal: Acta Physiologica (Oxford, England)

Article Title: Angiotensin‐(1–7) Alleviates Isoproterenol‐Induced Cardiac Hypertrophy by Suppressing Autophagy and Apoptosis Through the Synergistic Action of Mas Receptor and Angiotensin II Type 2 Receptor

doi: 10.1111/apha.70200

Figure Lengend Snippet: Physical and chemical properties and biosafety assessment of Ang‐(1–7). (A) Key molecular properties: molecular weight (MW), net charge, isoelectric point (pI), hydrophobicity (GRAVY index), and aromaticity. (B, C) Topological structure predicted by Deep TMHMM, showing a spherical soluble conformation (no transmembrane helices) and predicted functional sites (red‐intracellular; blue‐extracellular). (D) Hemolytic activity ( n = 6). (E) Cytotoxicity assessment in H9c2, HepG2, and NRK‐52E cell lines ( n = 6). Cell viability was determined using the Cell Counting Kit‐8 (CCK‐8) assay. The data is expressed as an mean ± standard deviation (SD).

Article Snippet: After 7 days' acclimatization, animals were randomized (blinded assessment) into five groups ( n = 6 per group) receiving daily Subcutaneous injection for 7 days [ , , ]:Ctrl: saline (Sanlian, Harbin, China); ISO: isoproterenol 5 mg/kg/day (Solarbio, II0200, Beijing, China; dissolved in DMSO); ISO + Ang‐(1–7): Ang‐(1–7) 576 μg/kg/day (MCE, HY‐12403, USA; in distilled water); ISO + Ang‐(1–7) + A‐779: A‐7791148 μg/kg/day (MCE, HY‐P0216, USA); ISO + Ang‐(1–7) + PD123319 : PD123319 5 mg/kg/day (MCE, HY‐10259A, USA); ISO + Ang‐(1–7) + PD123319 + A779.

Techniques: Molecular Weight, Functional Assay, Activity Assay, Cell Counting, CCK-8 Assay, Standard Deviation

Journal: Acta Physiologica (Oxford, England)

Article Title: Angiotensin‐(1–7) Alleviates Isoproterenol‐Induced Cardiac Hypertrophy by Suppressing Autophagy and Apoptosis Through the Synergistic Action of Mas Receptor and Angiotensin II Type 2 Receptor

doi: 10.1111/apha.70200

Figure Lengend Snippet: Effects of Ang‐(1–7) on isoproterenol (ISO)‐induced cardiac hypertrophy and fibrosis. (A) HE staining and Masson staining of myocardial sections (scale bar = 20 μm). (B, C) Quantitative analysis of myocardial cell cross‐sectional area and collagen volume fraction. (D, E) Macroscopic morphology and cross‐sectional observation of the heart, with measurement of heart weight/body weight ratio (HW/BW) (scale bar = 2 mm). (F) Western blot analysis of ANP, BNP, and β‐MHC expression. (G–I) Quantitative analysis of band intensity normalized to GAPDH. The data is expressed as an mean ± standard deviation (SD) ( n = 5). A + I, Ang‐(1–7) + ISO; AntM+A + I, A‐779 + Ang‐(1–7) + ISO; AntA+A + I, PD123319 + Ang‐(1–7) + ISO. *** p < 0.001 versus Ctrl; ### p < 0.001 versus ISO; &&& p < 0.001, && p < 0.01, & p < 0.05 versus Ang‐(1–7) + ISO.

Article Snippet: After 7 days' acclimatization, animals were randomized (blinded assessment) into five groups ( n = 6 per group) receiving daily Subcutaneous injection for 7 days [ , , ]:Ctrl: saline (Sanlian, Harbin, China); ISO: isoproterenol 5 mg/kg/day (Solarbio, II0200, Beijing, China; dissolved in DMSO); ISO + Ang‐(1–7): Ang‐(1–7) 576 μg/kg/day (MCE, HY‐12403, USA; in distilled water); ISO + Ang‐(1–7) + A‐779: A‐7791148 μg/kg/day (MCE, HY‐P0216, USA); ISO + Ang‐(1–7) + PD123319 : PD123319 5 mg/kg/day (MCE, HY‐10259A, USA); ISO + Ang‐(1–7) + PD123319 + A779.

Techniques: Staining, Western Blot, Expressing, Standard Deviation

Journal: Acta Physiologica (Oxford, England)

Article Title: Angiotensin‐(1–7) Alleviates Isoproterenol‐Induced Cardiac Hypertrophy by Suppressing Autophagy and Apoptosis Through the Synergistic Action of Mas Receptor and Angiotensin II Type 2 Receptor

doi: 10.1111/apha.70200

Figure Lengend Snippet: Ang‐(1–7) improves ISO‐induced ventricular remodeling and dysfunction via MasR and AT 2 R. (A) Representative M‐mode echocardiograms from each group (scale bar = 2 mm). (B, C) Quantification of left ventricular internal diameters at systole (LVIDs) and diastole (LVIDd). (D, E) Left ventricular ejection fraction (LVEF%) and fractional shortening (LVFS%). (F, G) Left ventricular posterior wall thickness at systole (LVPWs) and diastole (LVPWd). (H) Representative immunofluorescence images of cardiomyocytes (green, α‐actinin; blue, nuclei; scale bar = 10 μm). (I) Relative mRNA expression of ANP, BNP, and β‐MHC ( n = 6). The data is expressed as an mean ± standard deviation (SD) ( n = 5). A + I, Ang‐(1–7) + ISO; AntM + A + I, A‐779 + Ang‐(1–7) + ISO; AntA + A + I, PD123319 + Ang‐(1–7) + ISO; AntM + AntA + A + I, A‐779 + PD123319 + Ang‐(1–7) + ISO. *** p < 0.001, ** p < 0.01 versus Ctrl; ### p < 0.001, ## p < 0.01 versus ISO; &&& p < 0.001, && p < 0.01, & p < 0.05 versus Ang‐(1–7) + ISO, & p < 0.05 versus ISO.

Article Snippet: After 7 days' acclimatization, animals were randomized (blinded assessment) into five groups ( n = 6 per group) receiving daily Subcutaneous injection for 7 days [ , , ]:Ctrl: saline (Sanlian, Harbin, China); ISO: isoproterenol 5 mg/kg/day (Solarbio, II0200, Beijing, China; dissolved in DMSO); ISO + Ang‐(1–7): Ang‐(1–7) 576 μg/kg/day (MCE, HY‐12403, USA; in distilled water); ISO + Ang‐(1–7) + A‐779: A‐7791148 μg/kg/day (MCE, HY‐P0216, USA); ISO + Ang‐(1–7) + PD123319 : PD123319 5 mg/kg/day (MCE, HY‐10259A, USA); ISO + Ang‐(1–7) + PD123319 + A779.

Techniques: Immunofluorescence, Expressing, Standard Deviation

Journal: Acta Physiologica (Oxford, England)

Article Title: Angiotensin‐(1–7) Alleviates Isoproterenol‐Induced Cardiac Hypertrophy by Suppressing Autophagy and Apoptosis Through the Synergistic Action of Mas Receptor and Angiotensin II Type 2 Receptor

doi: 10.1111/apha.70200

Figure Lengend Snippet: Ang‐(1–7) regulates the expression of MasR and AT 2 R in H9c2 cardiomyocytes and exhibits cross‐inhibition with receptor antagonists. (A) Immunofluorescence images and immunofluorescence intensity of MasR (green), AT 2 R (red), and DAPI (blue) in H9c2 cells from different treatment groups, reflecting receptor expression levels (scale bar = 80 μm) ( n = 5). (B) Shows the expression of AT 2 R and MasR mRNA in H9c2 cells ( n = 6). (C) Analyzes MasR expression and relative thermal stability in cardiomyocytes at different temperatures (37°C–58°C). The data is expressed as an mean ± standard deviation (SD) ( n = 5). A + I, Ang‐(1–7) + ISO; AntM + A + I, A‐779 + Ang‐(1–7) + ISO; AntA + A + I, PD123319 + Ang‐(1–7) + ISO; AntM + AntA + A + I, A‐779 + PD123319 + Ang‐(1–7) + ISO. *** p < 0.001, * p < 0.05 versus Ctrl; #### p < 0.0001, ### p < 0.001 versus ISO; &&& p < 0.001, && p < 0.01, & p < 0.05 versus Ang‐(1–7) + ISO.

Article Snippet: After 7 days' acclimatization, animals were randomized (blinded assessment) into five groups ( n = 6 per group) receiving daily Subcutaneous injection for 7 days [ , , ]:Ctrl: saline (Sanlian, Harbin, China); ISO: isoproterenol 5 mg/kg/day (Solarbio, II0200, Beijing, China; dissolved in DMSO); ISO + Ang‐(1–7): Ang‐(1–7) 576 μg/kg/day (MCE, HY‐12403, USA; in distilled water); ISO + Ang‐(1–7) + A‐779: A‐7791148 μg/kg/day (MCE, HY‐P0216, USA); ISO + Ang‐(1–7) + PD123319 : PD123319 5 mg/kg/day (MCE, HY‐10259A, USA); ISO + Ang‐(1–7) + PD123319 + A779.

Techniques: Expressing, Inhibition, Immunofluorescence, Standard Deviation

Journal: Acta Physiologica (Oxford, England)

Article Title: Angiotensin‐(1–7) Alleviates Isoproterenol‐Induced Cardiac Hypertrophy by Suppressing Autophagy and Apoptosis Through the Synergistic Action of Mas Receptor and Angiotensin II Type 2 Receptor

doi: 10.1111/apha.70200

Figure Lengend Snippet: Ang‐(1–7) regulates the expression of MasR and AT 2 R and their heterodimer formation. (A) Immunofluorescence staining images of MasR (green), AT 2 R (red), and DAPI (blue) in myocardial tissue from mice in each treatment group. Arrows indicate the localization of receptors in the myocardium (scale bar = 20 μm) ( n = 5). (B) Protein expression and quantitative analysis of MasR and AT 2 R in myocardial tissue ( n = 6). (C) Molecular docking simulations showing the interactions between MasR and AT 2 R, as well as between Ang‐(1–7) and the receptors, along with the binding energies (ΔG) for each interaction. (D) Immunoprecipitation (Co‐IP): Interaction between MasR and AT 2 R in cardiac muscle tissue. The data is expressed as an mean ± standard deviation (SD). A + I, Ang‐(1–7) + ISO; AntM + A + I, A‐779 + Ang‐(1–7) + ISO; Ant A + A + I, PD123319 + Ang‐(1–7) + ISO. * p < 0.05 versus Ctrl; ### p < 0.001 versus ISO; &&& p < 0.001, & p < 0.05 versus Ang‐(1–7) + ISO.

Article Snippet: After 7 days' acclimatization, animals were randomized (blinded assessment) into five groups ( n = 6 per group) receiving daily Subcutaneous injection for 7 days [ , , ]:Ctrl: saline (Sanlian, Harbin, China); ISO: isoproterenol 5 mg/kg/day (Solarbio, II0200, Beijing, China; dissolved in DMSO); ISO + Ang‐(1–7): Ang‐(1–7) 576 μg/kg/day (MCE, HY‐12403, USA; in distilled water); ISO + Ang‐(1–7) + A‐779: A‐7791148 μg/kg/day (MCE, HY‐P0216, USA); ISO + Ang‐(1–7) + PD123319 : PD123319 5 mg/kg/day (MCE, HY‐10259A, USA); ISO + Ang‐(1–7) + PD123319 + A779.

Techniques: Expressing, Immunofluorescence, Staining, Binding Assay, Immunoprecipitation, Co-Immunoprecipitation Assay, Standard Deviation

Journal: Acta Physiologica (Oxford, England)

Article Title: Angiotensin‐(1–7) Alleviates Isoproterenol‐Induced Cardiac Hypertrophy by Suppressing Autophagy and Apoptosis Through the Synergistic Action of Mas Receptor and Angiotensin II Type 2 Receptor

doi: 10.1111/apha.70200

Figure Lengend Snippet: Ang‐(1–7) attenuates ISO‐induced excessive autophagy and apoptosis in vivo by regulating MasR and AT 2 R. (A) Transmission electron microscopy (TEM) images showing autophagosomes, lysosomes, and damaged mitochondria in cardiomyocytes; quantification of damaged mitochondria is shown (scale bar = 5 μm, 2 μm n = 4). (B) Western blot analysis of autophagy‐related proteins LC3‐II/I, Beclin1, and P62 ( n = 5). (C) Western blot analysis of apoptosis‐related proteins Bcl‐2, Bax, and cleaved caspase‐3 ( n = 5). (D, E) Quantitative densitometry analysis of autophagy‐ and apoptosis‐related proteins normalized to GAPDH. The data is expressed as an mean ± standard deviation (SD). A + I, Ang‐(1–7) + ISO; AntM + A + I, A‐779 + Ang‐(1–7) + ISO; Ant A + A + I, PD123319 + Ang‐(1–7) + ISO. *** p < 0.001 versus Ctrl; ### p < 0.001, # p < 0.05 versus ISO; &&& p < 0.001, && p < 0.01, & p < 0.05 versus Ang‐(1–7) + ISO.

Article Snippet: After 7 days' acclimatization, animals were randomized (blinded assessment) into five groups ( n = 6 per group) receiving daily Subcutaneous injection for 7 days [ , , ]:Ctrl: saline (Sanlian, Harbin, China); ISO: isoproterenol 5 mg/kg/day (Solarbio, II0200, Beijing, China; dissolved in DMSO); ISO + Ang‐(1–7): Ang‐(1–7) 576 μg/kg/day (MCE, HY‐12403, USA; in distilled water); ISO + Ang‐(1–7) + A‐779: A‐7791148 μg/kg/day (MCE, HY‐P0216, USA); ISO + Ang‐(1–7) + PD123319 : PD123319 5 mg/kg/day (MCE, HY‐10259A, USA); ISO + Ang‐(1–7) + PD123319 + A779.

Techniques: In Vivo, Transmission Assay, Electron Microscopy, Western Blot, Standard Deviation

Journal: Acta Physiologica (Oxford, England)

Article Title: Angiotensin‐(1–7) Alleviates Isoproterenol‐Induced Cardiac Hypertrophy by Suppressing Autophagy and Apoptosis Through the Synergistic Action of Mas Receptor and Angiotensin II Type 2 Receptor

doi: 10.1111/apha.70200

Figure Lengend Snippet: Ang‐(1–7) regulates autophagic flux and apoptosis in H9c2 cardiomyocytes through MasR and AT 2 R. (A) Representative images of TUNEL immunofluorescence staining (green fluorescence labels apoptotic cells) and Hoechst nuclear staining (blue) ( n = 6), with the Merge panel showing their overlay. (B) Western blot analysis of apoptosis related proteins (Bax, Bcl 2 , Cleaved caspase‐3) expression levels, with GAPDH as the internal control protein ( n = 3). (C) Western blot analysis of autophagy‐related proteins (LC3‐II, Beclin1, P62) expression levels, with GAPDH as the internal control protein ( n = 3). The data is expressed as an mean ± standard deviation (SD). A + I, Ang‐(1–7) + ISO; AntM + A + I, A‐779 + Ang‐(1–7) + ISO; Ant A + A + I, PD123319 + Ang‐(1–7) + ISO; AntM + AntA + A + I, A‐779 + PD123319 + Ang‐(1–7) + ISO.

Article Snippet: After 7 days' acclimatization, animals were randomized (blinded assessment) into five groups ( n = 6 per group) receiving daily Subcutaneous injection for 7 days [ , , ]:Ctrl: saline (Sanlian, Harbin, China); ISO: isoproterenol 5 mg/kg/day (Solarbio, II0200, Beijing, China; dissolved in DMSO); ISO + Ang‐(1–7): Ang‐(1–7) 576 μg/kg/day (MCE, HY‐12403, USA; in distilled water); ISO + Ang‐(1–7) + A‐779: A‐7791148 μg/kg/day (MCE, HY‐P0216, USA); ISO + Ang‐(1–7) + PD123319 : PD123319 5 mg/kg/day (MCE, HY‐10259A, USA); ISO + Ang‐(1–7) + PD123319 + A779.

Techniques: TUNEL Assay, Immunofluorescence, Staining, Fluorescence, Western Blot, Expressing, Control, Standard Deviation

Journal: Redox Biology

Article Title: Epithelial redox stress programs macrophage immunometabolism through a ZNF24-MIF–NF–κB pathway in chronic nonbacterial prostatitis

doi: 10.1016/j.redox.2026.104042

Figure Lengend Snippet: Pharmacological inhibition of MIF by ISO-1 attenuates prostatitis severity and suppresses M1 macrophage polarization in vivo . (A) Mechanical allodynia assessed by von Frey testing, showing reduced response frequency in ISO-1-treated mice. (B) Representative H&E staining of prostate sections showing decreased inflammatory infiltration after ISO-1 treatment. (C) Quantification of histopathological inflammation scores. (D) ELISA quantification of TNF-α, IL-6, and IL-1β in serum confirming cytokine downregulation. (E) Immunofluorescence staining for CD45 (red) indicating reduced immune cell infiltration. (F) Quantification of CD45 + immune cell infiltration in prostate tissues. (G–H) Flow cytometry of F4/80+CD11b + CD86 + macrophages showing reduced M1 subset after ISO-1. (I) Co-immunofluorescence for CD68 (red) and iNOS (green) showing reduced M1 macrophages in ISO-1-treated mice. (J) Western blot confirming downregulation of CD86 and iNOS in prostate tissues. Data are presented as the mean ± SD. ns, not significant; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. Abbreviations: CTR, control; EAP, experimental autoimmune prostatitis; qRT‒PCR, quantitative real-time PCR.

Article Snippet: Following the second immunization, the mice were treated daily for 14 days with ISO-1 (3.5 mg/kg in 10 % dimethyl sulfoxide (DMSO)/90 % corn oil; Cat. No. HY-16692; MCE) [ ] via intraperitoneal injection, whereas the controls received vehicle alone.

Techniques: Inhibition, In Vivo, Staining, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Flow Cytometry, Western Blot, Control, Real-time Polymerase Chain Reaction

Journal: Redox Biology

Article Title: Epithelial redox stress programs macrophage immunometabolism through a ZNF24-MIF–NF–κB pathway in chronic nonbacterial prostatitis

doi: 10.1016/j.redox.2026.104042

Figure Lengend Snippet: Epithelial ROS-ZNF24 axis drives MIF transcription and promotes CD74-dependent M1 macrophage polarization. (A–E) CD74 knockdown attenuates MIF-induced M1 macrophage polarization, as assessed by CD86 and iNOS expression (A–B), proinflammatory cytokine secretion (C), and flow cytometry analysis of F4/80 + CD86 + macrophages (D–E). (F–H) LPS stimulation induces MIF mRNA expression (F), protein expression (G), and secretion (H) in RWPE-1 prostate epithelial cells. (I–K) Transwell co-culture system showing that LPS-stimulated prostate epithelial cells promote M1 polarization and cytokine secretion in iBMDMs, which is suppressed by the MIF inhibitor ISO-1. (L–M) Increased epithelial oxidative stress in EAP mice and LPS-stimulated RWPE-1 cells, indicated by 8-OHdG staining (L) and intracellular ROS levels (M), respectively; NAC effectively reduces ROS accumulation. (N–O) ROS scavenging with NAC suppresses epithelial MIF expression and attenuates M1 marker expression in co-cultured macrophages. (P–T) ZNF24 is induced by epithelial ROS and directly regulates MIF transcription, as shown by ZNF24 expression (P), ZNF24 knockdown (Q), predicted ZNF24 binding motifs in the MIF promoter (R), and ChIP assays demonstrating enhanced ZNF24 binding upon LPS stimulation (S–T). Data are presented as mean ± SD. ns, not significant; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. Abbreviations: iBMDMs, immortalized bone marrow-derived macrophages; RWPE-1, human prostate epithelial cell line; siCD74, small interfering RNA targeting CD74; LPS, lipopolysaccharide; NAC, N-acetylcysteine; ROS, reactive oxygen species.

Article Snippet: Following the second immunization, the mice were treated daily for 14 days with ISO-1 (3.5 mg/kg in 10 % dimethyl sulfoxide (DMSO)/90 % corn oil; Cat. No. HY-16692; MCE) [ ] via intraperitoneal injection, whereas the controls received vehicle alone.

Techniques: Knockdown, Expressing, Flow Cytometry, Co-Culture Assay, Staining, Marker, Cell Culture, Binding Assay, Derivative Assay, Small Interfering RNA

Journal: Redox Biology

Article Title: Epithelial redox stress programs macrophage immunometabolism through a ZNF24-MIF–NF–κB pathway in chronic nonbacterial prostatitis

doi: 10.1016/j.redox.2026.104042

Figure Lengend Snippet: Pharmacological inhibition of NF-κB mitigates MIF-CD74-driven inflammation and M1 polarization in chronic prostatitis. (A) Heatmap of GSEA HALLMARK pathways showing increased inflammatory and NF-κB-related signaling in EAP, reversed by ISO-1. (B) Quantification of TNFA signaling via NF-κB scores from HALLMARK enrichment based on RNA-seq results derived from mouse model. (C) Heatmap of key NF-κB pathway genes, upregulated in EAP and suppressed by ISO-1 based on RNA-seq results derived from mouse model. (D–E) Single-cell transcriptomic analysis showing increased NF-κB pathway activity in EAP prostates using AddModuleScore. (F) IHC of p-p65 confirming NF-κB activation in EAP and inhibition by ISO-1 or CD74 blockade. (G) In vitro validation in iBMDMs: MIF-induced p65 phosphorylation reversed by si CD74 . (H) Confocal immunofluorescence showing MIF-induced nuclear translocation of p65 in iBMDMs, abolished by si CD74 . (I) Western blot showing suppression of MIF-induced CD86 and iNOS upregulation by the NF-κB inhibitor JSH-23. (J) Confocal staining showing decreased iNOS after JSH-23 treatment in MIF-stimulated iBMDMs. (K–L) Flow cytometry and quantification of F4/80+CD86 + macrophages showing reduced M1 polarization with JSH-23. (M) ELISA showing suppression of IL-6, IL-1β, and TNF-α secretion by JSH-23. Data are presented as the mean ± SD. ns, not significant; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. Abbreviations: GSEA, Gene Set Enrichment Analysis; HALLMARK, curated gene set collection; AddModuleScore, single-cell pathway activity scoring method; JSH-23, NF-κB inhibitor.

Article Snippet: Following the second immunization, the mice were treated daily for 14 days with ISO-1 (3.5 mg/kg in 10 % dimethyl sulfoxide (DMSO)/90 % corn oil; Cat. No. HY-16692; MCE) [ ] via intraperitoneal injection, whereas the controls received vehicle alone.

Techniques: Inhibition, RNA Sequencing, Derivative Assay, Activity Assay, Paraffin-embedded Immunohistochemistry, Activation Assay, In Vitro, Biomarker Discovery, Phospho-proteomics, Immunofluorescence, Translocation Assay, Western Blot, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Redox Biology

Article Title: Epithelial redox stress programs macrophage immunometabolism through a ZNF24-MIF–NF–κB pathway in chronic nonbacterial prostatitis

doi: 10.1016/j.redox.2026.104042

Figure Lengend Snippet: Schematic illustration of the epithelial ROS-ZNF24-MIF-CD74-PKM2-NF-κB signaling axis in chronic prostatitis. In response to epithelial injury or inflammatory stimuli, excessive ROS accumulate in prostatic epithelial cells, leading to the activation of the redox-responsive transcription factor ZNF24. Activated ZNF24 directly binds to the MIF promoter and drives MIF transcription, resulting in enhanced MIF production and release. Epithelial-derived MIF subsequently acts in a paracrine manner by binding to CD74 on macrophages. This interaction promotes the stabilization and nuclear translocation of PKM2, which acts as a co-activator to increase NF-κB signaling via p65 nuclear translocation. PKM2 activation also enhances glycolytic flux and contributes to mitochondrial dysfunction, further promoting M1 macrophage polarization and inflammatory cytokine production. Disruption of this axis via ISO-1, CD74 blockade, or PKM2 modulation (DASA-58) effectively mitigates chronic inflammation in the prostate.

Article Snippet: Following the second immunization, the mice were treated daily for 14 days with ISO-1 (3.5 mg/kg in 10 % dimethyl sulfoxide (DMSO)/90 % corn oil; Cat. No. HY-16692; MCE) [ ] via intraperitoneal injection, whereas the controls received vehicle alone.

Techniques: Activation Assay, Derivative Assay, Binding Assay, Translocation Assay, Disruption

Journal: bioRxiv

Article Title: Characterizing the SASP-Dependent Paracrine Spreading of Senescence Between Human Brain Cell Types

doi: 10.64898/2026.02.10.705129

Figure Lengend Snippet: A) Schematic depicting BulkSignalR pipeline which uses known ligand-receptor interactions and affected downstream pathways to analyze their activation based on our bulk RNAseq data from DMSO and BrdU treated human cell lines (created with BioRender). B) Venn diagram showing the number of receptors inferred from BulkSignalR to be activated across each of the five human cell types. Three receptors were identified in common between astrocytes (purple), endothelial cells (pink), and microglia (yellow) which were the cell types shown to be capable of receiving senescence signals and becoming SA β-gal positive: CXCR7, KREMEN2, and GIPR. Only CXCR7 was expressed in the cell types capable of entering secondary senescence (astrocytes, endothelial cells, microglia) ( , S3B). C) TPM expression values of CXCR7 , its ligand CXCL12 , and DPP4 which cleaves and inactivates CXCL12 in DMSO (grey) and BrdU (red) treated cell lines (n=3 replicates). D) Schematic of the four selected SASP inhibitors mechanisms of action: Bindarit is a CCL2 synthesis inhibitor which prevents p65 activation of the CCL2 gene at the promoter region, ISO-1 is a MIF antagonist, ACT-1004-1239 is a CXCR7 antagonist, and Sitagliptin inhibits DPP4 preventing its action of cleaving and inactivating CXCL12 (created with BioRender). Data was analyzed by two-way ANOVA with Tukey’s multiple comparisons test (C). All graphs show mean with error bars depicting standard deviation (ns, p>0.05, ** p<0.01, *** p<0.001).

Article Snippet: Treatment with migration inhibitor factor (MIF) antagonist ISO-1 (MedChemExpress, Catalog No. HY-16692) ( ) was utilized to prevent MIF-associated communication between senescence inducers and receiving cells.

Techniques: Activation Assay, RNA sequencing, Expressing, Standard Deviation

Journal: bioRxiv

Article Title: Characterizing the SASP-Dependent Paracrine Spreading of Senescence Between Human Brain Cell Types

doi: 10.64898/2026.02.10.705129

Figure Lengend Snippet: A) CCK8 viability assay in astrocytes showing normalized percent cell survival following treatment with DMSO control (grey) or DMSO + Bindarit (pink), DMSO + ISO-1 (blue), DMSO + ACT-1004-1239 (orange), or DMSO + Sitagliptin (green) at various concentrations (n=3 replicates). B) CCK8 viability assay in astrocytes showing normalized percent cell survival following treatment with 100 µM BrdU (red) or BrdU + Bindarit (pink), BrdU + ISO-1 (blue), BrdU + ACT-1004-1239 (orange), or BrdU + Sitagliptin (green) at various concentrations (n=3 replicates). C) CCK8 viability assay in microglia showing normalized percent cell survival following treatment with DMSO control (grey) or DMSO + Bindarit (pink), DMSO + ISO-1 (blue), DMSO + ACT-1004-1239 (orange), or DMSO + Sitagliptin (green) at various concentrations (n=3 replicates). D) CCK8 viability assay in microglia showing normalized percent cell survival following treatment with 100 µM BrdU (red) or BrdU + Bindarit (pink), BrdU + ISO-1 (blue), BrdU + ACT-1004-1239 (orange), or BrdU + Sitagliptin (green) at various concentrations (n=3 replicates). Data analyzed by one-way ANOVA with Dunnett’s correction for multiple comparisons (A-D). All graphs show mean with error bars depicting standard deviation (ns, p>0.05, * p<0.05, ** p<0.01, *** p<0.001).

Article Snippet: Treatment with migration inhibitor factor (MIF) antagonist ISO-1 (MedChemExpress, Catalog No. HY-16692) ( ) was utilized to prevent MIF-associated communication between senescence inducers and receiving cells.

Techniques: Viability Assay, Control, Standard Deviation

Journal: bioRxiv

Article Title: Characterizing the SASP-Dependent Paracrine Spreading of Senescence Between Human Brain Cell Types

doi: 10.64898/2026.02.10.705129

Figure Lengend Snippet: A) Quantification of percentage of SA β-gal positive astrocytes following 7-day treatment with 100 µM BrdU (red) along with 200 µM Bindarit (pink), 50 µM ISO-1 (blue), 200 µM ACT-1004-1239 (orange), or 2 µM Sitagliptin (green) (n=6 replicates). B) Quantification of percentage of SA β-gal positive microglia following 7-day treatment with 100 µM BrdU (red) along with 200 µM Bindarit (pink), 50 µM ISO-1 (blue), 200 µM ACT-1004-1239 (orange), or 2 µM Sitagliptin (green) (n=6 replicates). C) Timeline showing treatment with DMSO + Bindarit CM or BrdU + Bindarit CM for 7 days. Timeline showing treatment with DMSO or 100 µM BrdU along with SASP inhibitors (ISO-1, ACT-1004-1239, or Sitagliptin) for 7 days. Features of senescence were analyzed 8 days after the initial plating of cells (created with BioRender). D) Quantification of percentage of SA β-gal positive astrocytes following 7-day treatment with DMSO + Bindarit CM from astrocytes (grey), BrdU + Bindarit CM from astrocytes (red), DMSO CM from astrocytes + SASP inhibitor (grey), or BrdU CM from astrocytes + SASP inhibitor (red) (n=4 replicates). Quantification of percentage of SA β-gal positive astrocytes following 7-day treatment with DMSO CM from astrocytes + Bindarit (grey) or BrdU CM from astrocytes + Bindarit (red) (n=4 replicates). E) Quantification of percentage of SA β-gal positive astrocytes following 7-day treatment with DMSO + Bindarit CM from microglia (grey), BrdU + Bindarit CM from microglia (red), DMSO CM from microglia + SASP inhibitor (grey), or BrdU CM from microglia + SASP inhibitor (red) (n=4 replicates). Quantification of percentage of SA β-gal positive astrocytes following 7-day treatment with DMSO CM from microglia + Bindarit (grey) or BrdU CM from microglia + Bindarit (red) (n=4 replicates). F) Quantification of percentage of SA β-gal positive microglia following 7-day treatment with DMSO + Bindarit CM from microglia (grey), BrdU + Bindarit CM from microglia (red), DMSO CM from microglia + SASP inhibitor (grey), or BrdU CM from microglia + SASP inhibitor (red) (n=4 replicates). Quantification of percentage of SA β-gal positive astrocytes following 7-day treatment with DMSO CM from microglia + Bindarit (grey) or BrdU CM from microglia + Bindarit (red) (n=4 replicates). G) Quantification of percentage of SA β-gal positive microglia following 7-day treatment with DMSO + Bindarit CM from astrocytes (grey), BrdU + Bindarit CM from astrocytes (red), DMSO CM from astrocytes + SASP inhibitor (grey), or BrdU CM from astrocytes + SASP inhibitor (red) (n=4 replicates). Quantification of percentage of SA β-gal positive astrocytes following 7-day treatment with DMSO CM from astrocytes + Bindarit (grey) or BrdU CM from astrocytes + Bindarit (red) (n=4 replicates). Data analyzed by unpaired t-test (A-B) and two-way ANOVA with Tukey’s or Šídák’s multiple comparisons test (D-G). All graphs show mean with error bars depicting standard deviation (ns, p>0.05, * p<0.05, ** p<0.01, *** p<0.001).

Article Snippet: Treatment with migration inhibitor factor (MIF) antagonist ISO-1 (MedChemExpress, Catalog No. HY-16692) ( ) was utilized to prevent MIF-associated communication between senescence inducers and receiving cells.

Techniques: Standard Deviation