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Low concentrations of 17ß-estradiol protect single cardiac cells against metabolic stress-induced Ca²⁺ loading

Sofija Jovanovi, DVM, PhD^* , Aleksandar Jovanovi, MD, PhD^* , Win K. Shen, MD, FACC^* and Andre Terzic, MD, PhD^*

^* Division of Cardiovascular Diseases, Department of Internal Medicine, Mayo Clinic, Mayo Foundation, Rochester, Minnesota, USA
Tayside Institute of Child Health, Ninewells Hospital and Medical School, University of Dundee, Dundee, Scotland, United Kingdom

View larger version (13K): [in a new window]   Figure 1 Hypoxia-reoxygenation induces Ca2+ loading in both female and male cardiomyocytes. Time-course (left and central panels) of fluorescence in Fluo-3 loaded female and male cardiomyocytes exposed to hypoxia-reoxygenation, with average concentrations of intracellular Ca2+ prior and following hypoxia-reoxygenation (right panel). Bars represent mean ± SEM (n = 13–14). Hypoxia-reoxygenation was induced by application (3 min at 37°C), and removal of 2 mmol/liter DNP.

View larger version (42K): [in a new window]   Figure 2 The 17ß-estradiol (E2) prevents decreases hypoxia-reoxygenation–induced Ca2+ loading in female, but not male, cardiomyocytes. Epifluorescent digital images of Fluo-3AM loaded, and 10 nmol/liter pretreated cardiac cells from female (A) and male (B) hearts prior to and following hypoxia-reoxygenation (H/R). Horizontal bar corresponds to 30 µm. Time-course of averaged Fluo-3 fluorescence in cells presented in A (A1) and B (B1). AU: arbitrary units. (C) Average concentration of intracellular Ca2+ at rest, and following hypoxia-reoxygenation. Bars represent mean ± SEM (n = 6–7).

View larger version (54K): [in a new window]   Figure 3 Tamoxifen blocks the protective action of E2. Epifluorescent digital images of Fluo-3AM loaded cardiac cells from female hearts, pretreated with 10 nmol/liter E2 plus 1 µmol/liter tamoxifen, prior to and following hypoxia-reoxygenation (H/R). Horizontal bar corresponds to 30 µm. (A1) Time-course of averaged Fluo-3 fluorescence in cell presented in A. AU = arbitrary units. (B) Average concentration of intracellular Ca2+ at rest and following hypoxia-reoxygenation. Bars represent mean ± SEM (n = 5).

View larger version (42K): [in a new window]   Figure 4 17 Estradiol (E2) does not prevent hypoxia-reoxygenation–induced Ca2+ loading in female cardiomyocytes. Epifluorescent digital images of Fluo-3AM loaded cardiac cells from female hearts pretreated with 10 nmol/liter E2 prior to and following hypoxia-reoxygenation (H/R). Horizontal bar corresponds to 30 µm. (A1) Time-course of Fluo-3AM fluorescence in cell presented in A. AU: arbitrary units. (B) Average concentration of intracellular Ca2+ at rest and following hypoxia-reoxygenation. Bars represent mean ± SEM (n = 6).