Resveratrol inhibits cardiac hypertrophy via AMP-activated protein kinase and Akt - PubMed (original) (raw)

Resveratrol blunts phenylephrine-induced cardiac myocyte hypertrophy and protein synthesis via the p70S6K, eEF2, and calcineurin-NFAT signaling pathways. A, qualitative representation of the cell size of neonatal rat cardiac myocytes treated with ethanol (Control) or 50 μ

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resveratrol (Resv), in the absence or presence of 10 μ

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phenylephrine (PE) for 24 h. Changes in cell size were visualized following fixation on coverslips using mouse anti-α-actinin and Texas Red-conjugated donkey anti-mouse antibodies.B, measurement of protein synthesis using [3H]phenylalanine incorporation with counts expressed in DPM (disintegrations per minute). Values are means ± S.E. with each experiment performed in duplicate (n = 4). *, p < 0.01 versus Control; #, p < 0.05 versus PE; ^, p < 0.05 versus Resv; assessed by ANOVA and Bonferroni multiple comparisons test. C-E, representative immunoblot and densitometry of cellular extracts from cardiac myocytes treated as described above. Values are expressed as means ± S.E. and analyzed with the Kruskal-Wallis test followed by Dunn's multiple comparisons test. C, cell lysates blotted with anti-phospho-p70S6K (T389) and anti-actin antibodies; *, p < 0.05 versus Control; #,p < 0.01 versus PE (n = 9-10). D, cell lysates blotted with anti-phospho-p70S6K (T421/S424) and anti-actin antibodies; *, p < 0.05 versus Control; #,p < 0.05 versus PE (n = 5-6). E, cell lysates blotted with anti-phospho-eEF2 (T56) and anti-eEF2 antibodies;*, p < 0.05 versus Control; #, p < 0.01 versus PE (n = 8-9). F, cellular extracts of cardiac myocytes infected with Ad.NFAT-Luc-Promoter adenovirus and treated as described above were assessed for NFAT-dependent transcription, measured as luciferase activity expressed in relative light units. Values are means ± S.E. with each sample assessed in duplicate (n = 5);**, p < 0.001 versus Control; *,p < 0.05 versus Control; #, p < 0.001_versus_ PE, analyzed by ANOVA and Bonferroni multiple comparisons test. G and H, myocytes infected with Ad.NFAT-Luc-Promoter adenovirus were treated with 500 ng/ml (416 n

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) of CsA or 150 ng/ml (182 n

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) of 506 with or without 50 μ

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Resv, in the presence of 10 μ

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PE for 24 h. G, NFAT-dependent transcription measured from cellular extracts is presented as a percentage (%) compared with PE treatment, where values are means ± S.E. with each sample assessed in duplicate (n = 4); *, p < 0.001 versus Resv; #, p < 0.001 versus CsA; ^,p < 0.001 versus FK506, analyzed by ANOVA and Bonferroni multiple comparisons test. H, calcineurin activity measured from cellular extracts is presented as a percentage (%) compared with PE treatment, where values are means ± S.E. with each sample assessed in duplicate (n = 4-5), analyzed by ANOVA.