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Phenolic Metabolites Modulate Cardiomyocyte Beating in Response to Isoproterenol


Cardiovascular disease (CVD) is a public health concern, and the third cause of death worldwide. Several epidemiological studies and experimental approaches have demonstrated that consumption of polyphenol-enriched fruits and vegetables can promote cardioprotection. Thus, diet plays a key role in CVD development and/or prevention. Physiological β-adrenergic stimulation promotes beneficial inotropic effects by increasing heart rate, contractility and relaxation speed of cardiomyocytes. Nevertheless, chronic activation of β-adrenergic receptors can cause arrhythmias, oxidative stress and cell death. Herein the cardioprotective effect of human metabolites derived from polyphenols present in berries was assessed in cardiomyocytes, in response to chronic β-adrenergic stimulation, to disclose some of the underlying molecular mechanisms. Ventricular cardiomyocytes derived from neonate rats were treated with three human bioavailable phenolic metabolites found in circulating human plasma, following berries’ ingestion (catechol-O-sulphate, pyrogallol-O-sulphate, and 1-methylpyrogallol-O-sulphate). The experimental conditions mimic the physiological concentrations and circulating time of these metabolites in the human plasma (2 h). Cardiomyocytes were then challenged with the β-adrenergic agonist isoproterenol (ISO) for 24 h. The presence of phenolic metabolites limited ISO-induced mitochondrial oxidative stress. Likewise, phenolic metabolites increased cell beating rate and synchronized cardiomyocyte beating population, following prolonged β-adrenergic receptor activation. Finally, phenolic metabolites also prevented ISO-increased activation of PKA–cAMP pathway, modulating Ca2+ signalling and rescuing cells from an arrhythmogenic Ca2+ transients’ phenotype. Unexpected cardioprotective properties of the recently identified human-circulating berry-derived polyphenol metabolites were identified. These metabolites modulate cardiomyocyte beating and Ca2+ transients following β-adrenergic prolonged stimulation.

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β-Adrenergic receptors


Berry-derived polyphenols


Calcium calmodulin-dependent kinase II


Cyclic adenosine monophosphate


Cardiovascular diseases




cAMP-dependent protein kinase A


Reactive oxidative species


Ryanodine receptors


Sarcoplasmic reticulum


Sarcoplasmic reticulum calcium-ATPase


G protein-coupled receptors


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The authors would like to thank Pedro Sampaio, from Cilia Regulation and Disease lab, CEDOC, for technical support regarding the cardiomyocyte beating measurement.


The present work was supported by Fundação para a Ciência e Tecnologia (FCT, Portugal) [ANR-FCT/BEX-BCM/0001/2013], iNOVA4Health Unit (UID/Multi/04462/2013), the Agence National de la Recherche (France) [Grant ANR-13-ISV1-0001-01] and by the laBex LERMIT. Fundação para a Ciência e Tecnologia provided individual financial support to AG (SFRH/BD/103155/2014), HLAV (IF/00185/2012) and CNS (IF/01097/2013).

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Correspondence to Helena L. A. Vieira.

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Handling Editor: Shazina Saeed.

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Supplementary Figure 1 Schematic representation of the different phenolic metabolites used. The compounds were present in the human plasma in different concentrations: catechol-O-sulphate: 12 µM; pyrogallol-O-sulphate: 6 µM; and 1-methylpyrogallol-O-sulphate: 3 µM. (TIF 439 KB)


Supplementary Figure 2 Cell Viability. Cell viability was detected using propidium iodide, of differentiated H9c2 cells, treated with phenolic metabolites for 2 h and exposed to ISO for 48 h. Data are mean ± SD. ****P < 0.0001 versus Control and ####P < 0.0001 versus ISO (ISO: Isoproterenol). (TIF 482 KB)

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Dias-Pedroso, D., Guerra, J., Gomes, A. et al. Phenolic Metabolites Modulate Cardiomyocyte Beating in Response to Isoproterenol. Cardiovasc Toxicol 19, 156–167 (2019).

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  • Cardiomyocytes
  • β-Adrenergic receptors
  • Isoproterenol
  • Human bioavailable phenolic metabolites
  • Polyphenols