E-2 beta stimulates ovine uterine artery endothelial cell H2S production in vitro by estrogen receptor-dependent upregulation of cystathionine beta-synthase and cystathionine gamma-lyase expression

摘要

Endogenous hydrogen sulfide (H2S) is a potent vasodilator and proangiogenic second messenger synthesized from L-cysteine by cystathionine beta-synthase (CBS) and cystathionine gamma-lyase (CTH). Estrogens are potent vasodilators that stimulate H2S biosynthesis in uterine arteries (UA) in vivo; however, the underlying mechanisms are unknown. We hypothesized that estrogens stimulate H2S biosynthesis in UA endothelial cells (UAEC) via specific estrogen receptor (ER)-dependent mechanisms. In cultured primary UAEC, treatment with estradiol-17 beta (E-2 beta) stimulated CBS and CTH mRNAs and proteins in a time- and concentration-dependent fashion. As little as 0.1 nM E-2 beta was effective in increasing CBS and CTH expressions and these stimulatory effects maximized with 10-100 nM E-2 beta at 48-72 h. E-2 beta also activated CBS and CTH promoters in UAEC, leading to CBS and CTH expression. Treatment with E-2 beta stimulated H2S production, which was blocked by specific inhibitors of either CBS or CTH and their combination and the ER antagonist ICI 182780. Treatment with either specific agonist of ER alpha or ER beta stimulated both CBS and CTH mRNA and protein expressions and H2S production to levels similar to that of E-2 beta. Specific antagonist of either ER alpha or ER beta blocked E-2 beta-stimulated CBS and CTH mRNA and protein expressions and H2S production. Combinations of either ER alpha or ER beta agonists or their antagonists had no additive effects. Thus, E-2 beta stimulates H2S production by upregulating CBS and CTH mRNA and protein expressions through specific ER alpha or ER beta-dependent CBS and CTH transcription in UAEC in vitro.