Cellular actions of testosterone in vascular cells: Mechanism independent of aromatization to estradiol

Campelo, Adrián Esteban; Cutini, P.; Massheimer, Virginia

doi:10.1016/j.steroids.2012.05.008

Cited by 49 publications

(30 citation statements)

References 48 publications

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“…Moreover, the vasodilatory effects of androgens have been proposed to be mediated by ion channel modulation in the smooth muscle cells, sidestepping classical nuclear receptor action by the AR [66]. However, although some studies have elicited that protection within the vasculature is due to non-genomic effects [67], others have demonstrated slower transcriptional effects mediated through the AR [68]. Expression of the AR has been found in most cells of the vasculature, including in platelets and endothelial cells [69].…”

Section: Discussionmentioning

confidence: 99%

Regulated expression of the prostacyclin receptor (IP) gene by androgens within the vasculature: Combined role for androgens and serum cholesterol

Eivers

Kinsella

2016

Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanism

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The prostanoid prostacyclin plays a key cardioprotective role within the vasculature. There is increasing evidence that androgens may also confer cardioprotection but through unknown mechanisms. This study investigated whether the androgen dihydrotestosterone (DHT) may regulate expression of the prostacyclin/I prostanoid receptor or, in short, the IP in platelet-progenitor megakaryoblastic and vascular endothelial cells. DHT significantly increased IP mRNA and protein expression, IP-induced cAMP generation and promoter (PrmIP)-directed gene expression in all cell types examined. The androgen-responsive region was localized to a cis-acting androgen response element (ARE), which lies in close proximity to a functional sterol response element (SRE) within the core promoter. In normal serum conditions, DHT increased IP expression through classic androgen receptor (AR) binding to the functional ARE within the PrmIP. However, under conditions of low-cholesterol, DHT led to further increases in IP expression through an indirect mechanism involving AR-dependent upregulation of SCAP expression and enhanced SREBP1 processing & binding to the SRE within the PrmIP. Chromatin immunoprecipitation assays confirmed DHTinduced AR binding to the ARE in vivo in cells cultured in normal serum while, in conditions of low cholesterol, DHT led to increased AR and SREBP1 binding to the functional ARE and SRE cis-acting elements, respectively, within the core PrmIP resulting in further increases in IP expression. Collectively, these data establish that the human IP gene is under the transcriptional regulation of DHT, where this regulation is further influenced by serum-cholesterol levels. This may explain, in part, some of the protective actions of androgens within the vasculature. Acknowledgements:We thank Dr Helen M. Reid for reading the manuscript during its preparation.

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Section: Discussionmentioning

confidence: 99%

Regulated expression of the prostacyclin receptor (IP) gene by androgens within the vasculature: Combined role for androgens and serum cholesterol

Eivers

Kinsella

2016

Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanism

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“…In order to establish a pure EC culture, after 3 days of culture, the ring explants were removed and ECs were allowed to reach confluency. The identity of the ECs was determined i) by phase-contrast microscope observation of the characteristic morphology of cobblestone shape growth in confluent monolayer, ii) by positive immunocytochemistry reactivity to Factor VIII and to anti-Vimentin, clone V9 using the DakoCytomation EnVision system, and iii) by the bioability to synthesize NO (Campelo et al 2012). Cells from passages 2-7 were used for all experiments.…”

Section: Ec Culturementioning

confidence: 99%

“…Platelet isolation For platelet aggregation assays, rat platelet-rich plasma (PRP) was obtained as described previously (Cutini et al 2012). For platelet adhesion assays, PRP was fixed in 4% (v/v) formaldehyde/PBS for 10 min, suspended in PBS (pH 7.4) at room temperature, and centrifuged at 750 g for 10 min.…”

Section: Platelet Assaysmentioning

confidence: 99%

“…Platelet aggregation assay Platelet aggregation was measured using a turbidimetric assay as described previously (Cutini et al 2012). ECs were seeded on 24-multiwell culture plates (NUNC) at a density of 3!10 4 cells/well in DMEM supplemented with 10% (v/v) FCS and allowed to grow to 60-70% of confluency.…”

Section: Platelet Assaysmentioning

confidence: 99%

“…When specific antagonist/inhibitors were used, they were added 30 min before steroid treatment. Nitrites (NO 2 K ) were measured in the incubation media as a stable and non-volatile breakdown product of the NO released, employing the spectrometric Griess reaction (Campelo et al 2012). Briefly, once treatment is finished, aliquots of culture medium supernatant were mixed with Griess reagent (1% sulphanilamide and 0.1% naphthylene diamine dihydrochloride in 2.5% phosphoric acid) and incubated 10 min at room temperature.…”

Section: Measurement Of No Productionmentioning

confidence: 99%

See 2 more Smart Citations

Differential regulation of endothelium behavior by progesterone and medroxyprogesterone acetate

Cutini¹,

Campelo²,

Massheimer³

2013

Journal of Endocrinology

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Medroxyprogesterone acetate (MPA) is a synthetic progestin commonly used in hormone replacement therapy (HRT). The aim of this research was to study and compare the effect of progesterone (Pg) and MPA on the regulation of cellular events associated with vascular homeostasis and disease. Platelet adhesion to endothelial cells (ECs), nitric oxide (NO) production, and cell migration were studied using murine ECs in vitro exposed to the progestins. After 7 min of treatment, MPA significantly inhibited NO synthesis with respect to control values; meanwhile, Pg markedly increased vasoactive production. In senile ECs, the stimulatory action of Pg decreases; meanwhile, MPA maintained its ability to inhibit NO synthesis. The presence of RU486 antagonized the action of each steroid. When ECs were preincubated with PD98059 (MAPK inhibitor) or chelerythrine (protein kinase C (PKC) inhibitor) before Pg or MPA treatment, the former totally suppressed the steroid action, but the PKC antagonist did not affect NO production. In the presence of a PI3K inhibitor (LY294002), a partial reduction in Pg effect and a reversal of MPA action were detected. Using indomethacin, the contribution of the cyclooxygenase (COX) pathway was also detected. On platelet adhesion assays, Pg inhibited and MPA stimulated platelet adhesion to ECs. Under inflammatory conditions, Pg prevented platelet adhesion induced by lipopolysaccharide (LPS); meanwhile, MPA potentiated the stimulatory action of LPS. Finally, although both steroids enhanced migration of ECs, MPA exhibited a greater effect. In conclusion, the data presented in this research provide evidence of a differential regulation of vascular function by Pg and MPA.

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Endocrine Disruptors Differently Influence Estrogen Receptor β and Androgen Receptor in Male and Female Rat VSMC

Pellegrini

Bulzomi

Lecis

et al. 2014

Journal Cellular Physiology

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Sex steroid hormones differently control the major physiological processes in male and female organisms. In particular, their effects on vascular smooth muscle cells (VSMCs) migration are at the root of sex/gender-related differences reported in the cardiovascular system. Several exogenous substances, defined endocrine disruptor chemicals (EDCs), could interfere with these androgen and estrogen effects; however, the sex/gender-related susceptibility of VSMC motility to EDCs is completely unknown. Here, the effect of naturally occurring (naringenin, Nar) and synthetic (bisphenol A, BPA) EDCs on male and female VSMC motility has been evaluated. 17β-estradiol (E2, 0.1 nM-1 µM) induced a dose-dependent inhibition of motility in female-derived VSMC. In contrast, neither dihydrotestosterone (DHT, 0.01-100 nM) nor the common precursor of sex steroid hormones, testosterone (Tes, 0.01-100 nM) modified male-derived VSMC motility. Estrogen receptor (ER) β subtype-dependent activation of p38 was necessary for the E2 effect on cell motility. High BPA concentration prevented E2 effects in female-derived cells being without any effect in male-derived cells. Nar mimicked E2 effects on female-derived cells even in the presence of E2 or BPA. Intriguingly, Nar also inhibited the male-derived VSMC mobility. This latter effect was prevented by ERβ inhibitor, but not by the androgen receptor (AR) inhibitor. As a whole, ERβ-dependent signals in VSMC results more susceptible to the impact of EDCs than AR signals suggesting a possible high and overall susceptibility of female to EDCs. However, several male-derived cells, including VSMC, express ERβ, which could also serve as target of EDC disruption in male organisms.

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Cellular actions of testosterone in vascular cells: Mechanism independent of aromatization to estradiol

Cited by 49 publications

References 48 publications

Regulated expression of the prostacyclin receptor (IP) gene by androgens within the vasculature: Combined role for androgens and serum cholesterol

Regulated expression of the prostacyclin receptor (IP) gene by androgens within the vasculature: Combined role for androgens and serum cholesterol

Differential regulation of endothelium behavior by progesterone and medroxyprogesterone acetate

Endocrine Disruptors Differently Influence Estrogen Receptor β and Androgen Receptor in Male and Female Rat VSMC

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