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Cardiovascular Research 2002 56(1):43-51; doi:10.1016/S0008-6363(02)00505-9
© 2002 by European Society of Cardiology
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Copyright © 2002, European Society of Cardiology

Expression of estrogen receptor subtypes and neuronal nitric oxide synthase in neutrophils from women and men

Regulation by estrogen

Laura Moleroa, Margarita García-Durána, Joaquin Diaz-Recasensb, Luis Ricoa, Santos Casadoa and Antonio López-Farréa,*

aCardiovascular Research Laboratory, Fundación Jiménez Díaz, Avda. Reyes Catolicos 2, Madrid 28040, Spain
bObstetrics and Gynecology Department, Fundación Jiménez Díaz, Madrid, Spain

* Corresponding author. Tel.: +34-91-550-4821; fax: +34-91-549-4764 alopeza{at}fjd.es

Objectives: (a) To identify the subtype of estrogen receptor (ER) expressed in neutrophils from premenopausal women and in neutrophils from men under different estrogen conditions and (b) to analyze the association between the modifications in the expression of ER subtypes and neuronal nitric oxide synthase (nNOS) expression induced by estrogen. Methods: Neutrophils were isolated from pre-menopausal women during different stages of the menstrual cycle and from ten men for in vitro estrogen incubations. Results: Neutrophils from premenopausal women expressed both ER{alpha} and ERβ subtypes which were increased in the ovulatory phase of the menstrual cycle. Neutrophils derived from men also expressed ER{alpha} and ERβ but only ER{alpha} expression was enhanced by in vitro incubation with 17β-estradiol (10–8 mol/l). In vitro incubation of neutrophils from women with 17β-estradiol enhanced expression of both ER-{alpha} and ER-β subtypes. nNOS protein was overexpressed in neutrophils from premenopausal women during the ovulatory phase. 17β-Estradiol (10–8 mol/l) also increased nNOS protein expression in neutrophils derived from men. Mithramycin A (10–6 mol/l) and curcumin (10–6 mol/l), prevented the upregulation of nNOS and ER{alpha} in neutrophils derived from men, suggesting the involvement of AP-1 and Sp-1 transcription factors. Conclusions: Although the in vivo levels of circulating estrogen concentrations seem to be associated with overexpression of both ER{alpha} and ERβ in neutrophils from premenopausal women, which was further confirmed by the in vitro experiments with neutrophils from women, in vitro incubation of neutrophils from men with 17β-estradiol only increased ER{alpha} protein expression which was associated with enhanced expression of nNOS protein.

KEYWORDS Gender; Gene expression; Leukocyte; Nitric Oxide; Receptors


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