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The genomic action of 17 β‐estradiol accounts for altering calcium homeostasis in human endothelial cells
Author(s) -
Thor Der,
Rahimian Roshanak
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.626.24
Subject(s) - homeostasis , thapsigargin , endoplasmic reticulum , calcium , chemistry , extracellular , calcium metabolism , nitric oxide , fura 2 , microbiology and biotechnology , medicine , endocrinology , biology , biochemistry , enzyme , organic chemistry , cytosol
Our preliminary studies showed 17 β‐estradiol (E2)‐regulated Ca 2+ homeostasis may play a role in enhancing nitric oxide production in human endothelial cells, EA.hy926. Here, we investigated the genomic and nongenomic actions of E2 on intracellular calcium concentration ([Ca 2+ ] i ) in endothelial cells. Cells were treated with either a) E2 (1 μM) in the presence or absence of the transcription inhibitor actinomycin D (1 μg/mL) for 24 h or b) E2 (1 μM) for 5 minutes prior to the analysis of [Ca 2+ ] i . Using a spectrofluorometer, [Ca 2+ ] i from cells loaded with the Ca 2+ ‐sensitive dye, Fura 2‐AM, was monitored in the absence of extracellular Ca 2+ . The sarco/endoplasmic reticulum Ca 2+ ATPase inhibitor, thapsigargin (TG, 1 μM), was used to induce Ca 2+ release from the endoplasmic reticulum. TG‐induced [Ca 2+ ] i increase from cells treated with actinomycin D plus E2 was not significantly different from cells treated with vehicle (ethanol). However, the extent of TG‐induced [Ca 2+ ] i increase was significantly lower in vehicle‐ or actinomycin D‐ plus E2‐treated cells as compared with that observed in E2‐treated cells, indicating that gene transcription is required for E2‐mediated alteration in Ca 2+ response. Five minutes of E2 treatment did not alter TG‐induced [Ca 2+ ] i increase as compared to vehicle‐treated cells. Our results suggest that E2 regulates Ca 2+ homeostasis in EA.hy926 via a genomic mechanism. Supported by NHLBI.