Mechanistic analysis of resveratrol in cardiac hypertrophy by network pharmacology and animal experiments

Figures

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  Figure 1.. Flowchart of our research. TCMSP, traditional Chinese medicine systems pharmacology database and analysis platform; GEO, Gene Expression Omnibus; HC, hypertrophic cardiomyopathy; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; PPI, protein-protein interaction.

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  Figure 2.. Target characteristics of resveratrol for HC. (A) Molecular structure of resveratrol. (B) A total of 129 differential gene expression profiles for male HC, demonstrating 43 up- and 86 downregulated genes. (C) A total of 215 differential gene expression profiles in female HC, demonstrating 140 up- and 75 downregulated genes. (D) Common targets of resveratrol and HC illustrated using a Venn diagram. HC, hypertrophic cardiomyopathy; CDKN1A, cyclin-dependent kinase inhibitor 1A; ALOX5, arachidonate 5-lipoxygenase; APP, amyloid β precursor protein; CA3, carbonic anhydrase 3; FOS, Fos proto-oncogene; NFKBIA, NFκB inhibitor α.

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  Figure 3.. Molecular docking model and biological functions of targets. (A) Molecular docking model and interaction residues of the binding of Res to STAT3 (docking score, −4.9; interaction residues, Glu612 and Ser613). (B) Enrichment analysis of 8 overlapping targets. (C) Association of pathways regulated by Res with apoptosis and/or autophagy. Res, resveratrol; PDGF, platelet-derived growth factor; SARS, severe acute respiratory syndrome; FDR, false discovery rate.

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  Figure 4.. Network of protein-protein interactions of Res on hypertrophic cardiomyopathy with screening of core proteins. (A) Protein interaction network of Res in male patients with hypertrophic cardiomyopathy generated using the BisoGenet 1.0 plugin for Cytoscape 3.9, with node area and font size positively associated with protein degree value. (B) With regard to the effect of Res in male patients with HC, proteins encoded by core genes were screened using the CytoScape 3.9 MCODE plugin, with node area, font size and color intensity positively associated with the core protein degree values. (C) Protein interaction network of Res in female patients with hypertrophic cardiomyopathy generated using the BisoGenet 1.0 plugin for Cytoscape 3.9, with node area and font size positively correlated with protein degree value. (D) With regard to the effect of Res in female patients with HC, proteins encoded by core genes were screened using the CytoScape 3.9 MCODE plugin, with node area, font size and color intensity positively associated with the core protein degree values. Res, resveratrol.

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  Figure 5.. Res reverses ISO-induced myocardial hypertrophy. (A) HW/BW and (B) LVW/BW ratio. (C) mRNA expression levels of ANP were detected using reverse transcription-quantitative PCR. (D) Protein expression levels of ANP were detected using western blotting and normalized to GAPDH. *P<0.05 and **P<0.01 vs. control; #P<0.05 and ##P<0.01 vs. ISO; &P<0.05 and &&P<0.01 vs. Res. ISO, isoprenaline; Res, resveratrol; 3-MA, 3-methyladenine; HW, heart weight; BW, body weight; LVW, left ventricle weight; ANP, atrial natriuretic peptide.

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  Figure 6.. Res reverses ISO-induced cardiac dysfunction. Echocardiographic images of the left ventricle by the short-axis view and electrocardiography findings. (A) Representative M-mode echocardiograms in different groups. (B) EF and (C) FS were decreased in ISO group and were increased in Res group. (D) LVESd and (E) LVEDd were increased in ISO group and were decreased in Res group and LVESd was increased in Res+3-MA group. **P<0.01 vs. control; #P<0.05 and ##P<0.01 vs. ISO; &&P<0.01 vs. Res. EF, ejection fraction; FS, fractional shortening; LVESd, left ventricular end systolic diameter; LVEDd, left ventricular end diastolic diameter; ISO, isoprenaline; Res, resveratrol; 3-MA, 3-methyladenine.

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  Figure 7.. Res decreases ISO-induced oxidative stress of heart. (A) MDA and (B) LDH levels in heart tissues. **P<0.01 vs. control; #P<0.05 and ##P<0.01 vs. ISO; &P<0.05 vs. Res. MDA, malondialdehyde; LDH, lactate dehydrogenase; ISO, isoprenaline; Res, resveratrol; 3-MA, 3-methyladenine.

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  Figure 8.. Res downregulates the apoptosis pathway and upregulates the autophagy pathway. (A) Protein expression levels of CytC, Bcl-2 and Bax. Semi-quantification of the protein expression levels of (B) CytC, (C) Bcl-2 (arrow) and (D) Bax normalized to GAPDH protein expression. (E) Protein expression levels of Atg5. *P<0.05 and **P<0.01 vs. control; #P<0.05 and ##P<0.01 vs. ISO; &P<0.05 and &&P<0.01 vs. Res. CytC, cytochrome C; ISO, isoprenaline; Res, resveratrol; 3-MA, 3-methyladenine; Atg5, autophagy-related gene 5.