Divergent Effects of Resveratrol on Rat Cardiac Fibroblasts and Cardiomyocytes.

Garret Degagne,Jeffrey T Wigle,Sampath Krishnan,Thomas Netticadan,Xavier Lieben Louis,Rebecca Cummer,Laura Chan,Zach Meikle,Liping Yu,Shannon Meikle

doi:10.3390/molecules24142604

Garret Degagne, Jeffrey T Wigle + Show 8 more

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https://doi.org/10.3390/molecules24142604

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Abstract

In this study, we tested the potential cardioprotective effects of the phytoalexin resveratrol (Rsv) on primary adult rat cardiac fibroblasts (CF), myofibroblasts (MF) and cardiomyocytes. Adult rat CF and cardiomyocytes were isolated from male 10-week old Sprague–Dawley rats, cultured for either 24 h (cardiomyocytes) or 48 h (CF) before treatments. To isolate MF, CF were trypsinized after 48 h in culture, seeded in fresh plates and cultured for 24 h prior to treatment. All three cells were then treated for a further 24 h with a range of Rsv doses. In CF and MF, cell proliferation, viability, apoptosis assays were performed with or without Rsv treatment for 24 h. In cardiomyocytes, cell viability and apoptosis assay were performed 24 h after treatment. In separate experiments, CF was pre-incubated with estrogen, tamoxifen and fulvestrant for 30 min prior to Rsv treatment. Rsv treatment decreased proliferation of both fibroblasts and myofibroblasts. Rsv treatment also increased the proportion of dead CF and MF in a dose dependent manner. However, treatment with Rsv did not induce cell death in adult cardiomyocytes. There was an increase in the percentage of cells with condensed nuclei with Rsv treatment in both CF and MF, but not in cardiomyocytes. Treatment with estrogen, tamoxifen and fulvestrant alone or in combination with Rsv did not have any additional effects on CF survival. Our results demonstrate that treatment with Rsv can inhibit cell proliferation and induce cell death in rat CF and MF, while not affecting cardiomyocyte survival. We also demonstrated that the induction of cell death in CF with Rsv treatment was independent of estrogen receptor alpha (ERα) signaling.

Highlights

Heart failure (HF) is the end stage clinical manifestation of heart disease [1]
Cell count analyses of red and green stained cells representing live and dead cells, respectively, showed that Rsv treatment increased the percentage of dead cells in both cardiac fibroblasts (CF) and MF in a dose dependent manner (Figure 1A–C)
Rsv treatment did not alter the rate of cardiomyocyte cell death (Figure 1C)

Summary

Introduction

Heart failure (HF) is the end stage clinical manifestation of heart disease [1]. HF is a growing epidemic in Canada, with 50,000 new cases diagnosed each year [2]. Cardiac fibroblasts and cardiomyocytes are two cell types that play major roles in normal cardiac function, as well as in HF [3,4,5]. During a myocardial infarction (MI), cardiomyocytes die due to a lack of blood supply [6]. After cardiomyocyte death and the subsequent inflammatory response to clear damaged cells and debris, resident cardiac fibroblasts (CF) are activated and initiate the wound healing process [7].

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