Estrogen metabolites in the release of inflammatory mediators from human amnion-derived cells

AimsHuman amnion-derived cells have been used as in vitro models to test the release of inflammatory mediators, such as arachidonic acid (AA) and prostaglandin E₂ (PGE₂). We compared estrogen metabolites for their ability to induce AA release, to influence PGE₂ production and to interact toward intracellular estrogen receptors (ERs).Main methodsMetabolite effects on AA and PGE₂ release were examined by radiolabelled substrate incorporation and by colorimetric enzyme immunoassays, respectively. [³H]17-β-estradiol binding displacements were performed on Ro-20-1724 treated whole cells.Key findingsIn WISH cells, estrone, 2-hydroxy-estrone and estriol induced a rapid dose dependent release of AA that was not inhibited by cycloheximide. Estrone and 2-hydroxy-estrone showed biphasic dose–response curves of PGE₂, whereas estriol and 16-α-hydroxy-estrone increased PGE₂ levels at high concentrations. 2-methoxy-estrone, 4-hydroxy-estradiol and 4-hydroxy-estrone did not significantly affect PGE₂ release. 2-methoxy-estradiol and 2-hydroxy-estradiol decreased the PGE₂ release. Effects of metabolites on PGE₂ were inhibited by cycloheximide and by the ER antagonist tamoxifen. In AV3 cells PGE₂ production was poorly detectable. On Ro-20-1724 treated WISH cells the K_i of 17-β-estradiol was 29.2 ± 5.4 nM. Estrone, 2-methoxy-estrone and 2-methoxy-estradiol showed similar affinity values. The hydroxyl substituent at position 2, 4 and 16 decreased or markedly increased the affinity for estradiol or estrone derivatives, respectively.SignificanceThe estrogen metabolites induced nongenomic effects on AA release from WISH cells. The influence on PGE₂ release was detectable only on WISH cells. These effects appeared genomic and mediated by intracellular ERs, whose properties seemed strongly dependent on intracellular cAMP levels.