Session: SAT 358-380-Steroid Hormone Biosynthesis & Metabolism
Poster Board SAT-368
In our experimental setup primary human skin fibroblasts were incubated for 1 to 14 days with different concentrations of testosterone (0.1 - 20 µM) as substrate for the aromatase. The estradiol concentration in the cell free culture medium was measured as analytical parameter of aromatase activity using the Roche Elecsys 2010 system. It could be shown that the measured estradiol concentration increases in human skin cells in a time- and concentration-dependent manner.
To determine a suitable aromatase inhibitor, the skin fibroblasts were treated for 3 days with 20 µM testosterone and different concentrations (1 - 50 µM) of apigenin, 7-OH-flavone and chrysin, respectively. In addition to the estradiol measurement the cell proliferation was determined by BrdU incorporation and the cytotoxicity by LDH assay. Our results reveal that both apigenin and chrysin are toxic to human skin cells at higher concentrations whereas 7-OH-flavone inhibits aromatase without toxic effects.
To study the stimulating effect on the aromatase activity, the dermal cells were treated for 3 days with 20 µM testosterone and different concentrations of the glucocorticoid dexamethasone (0.001 - 1 µM). Our data demonstrate that the aromatase activity of skin fibroblasts is stimulated 70-fold by dexamethasone at concentrations above 0.05 µM.
From these results we conclude that the described non radioactive assay is suitable to monitor aromatase activity in primary human skin cells.
Nothing to Disclose: HS, NZ, MB, RL, RK, SK, AB
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