17β-oestradiol acts as a negative modulator of insulin-induced lactotroph cell proliferation through oestrogen receptor α, via nitric oxide/guanylyl cyclase/cGMP.
OBJECTIVE
17β-oestradiol interacts with growth factors to modulate lactotroph cell population. However, contribution of isoforms of the oestrogen receptor in these activities is not fully understood. In the present study, we have established participation of α and β oestrogen receptors in effects of 17β-oestradiol on lactotroph proliferation induced by insulin and shown involvement of the NO/sGC/cGMP pathway.


METHODS
Cell cultures were prepared from anterior pituitaries of female rats to evaluate lactotroph cell proliferation using bromodeoxyuridine (BrdUrd) detection, protein expression by western blotting and cGMP by enzyme immunoassay.


RESULTS
In serum-free conditions, 17β-oestradiol and α and β oestrogen receptor agonists (PPT and DPN) failed to increase numbers of lactotroph cells undergoing mitosis. Co-incubation of 17β-oestradiol/insulin and PPT/insulin significantly decreased lactotroph mitogenic activity promoted by insulin alone. Both ICI 182780 and NOS inhibitors (L-NMMA and L-NAME) induced reversal of the anti-proliferative effect promoted by 17β-oestradiol/insulin and PPT/insulin. Moreover, 17β-oestradiol, PPT and insulin increased sGC α1 protein expression and inhibited β1, whereas co-incubation of 17β-oestradiol/insulin or PPT/insulin induced increases of the two isoforms α1 and β1. 17β-oestradiol and insulin reduced cGMP production, while 17β-oestradiol/insulin co-incubation increased this cyclic nucleotide.


CONCLUSIONS
Our results suggest that 17β-oestradiol is capable of arresting lactotroph proliferation induced by insulin through ER α with participation of the signalling NO/sGC/cGMP pathway.
Gutiérrez
S
S
Center of Electron Microscopy, Faculty of Medical Sciences, National University of Córdoba, Córdoba, Argentina. silvina@cmefcm.uncor.edu
Petiti
J P
JP
Sosa
L d V
Ld
Fozzatti
L
L
De Paul
A L
AL
Masini-Repiso
A M
AM
Torres
A I
AI
D000818:Animals; D049109:Cell Proliferation; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000378:metabolism; D004958:Estradiol / Q000378:metabolism*; D047628:Estrogen Receptor alpha / Q000378:metabolism*; D005260:Female; D006162:Guanylate Cyclase / Q000378:metabolism*; D007328:Insulin / Q000378:metabolism*; D052682:Lactotrophs / Q000166:cytology*; D009569:Nitric Oxide / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar
D047628:Estrogen Receptor alpha; D007328:Insulin; D009569:Nitric Oxide; D004958:Estradiol; D006162:Guanylate Cyclase; D006152:Cyclic GMP