Estrogen increases endothelial nitric oxide synthase via estrogen receptors in rat cerebral blood vessels: effect preserved after concurrent treatment with medroxyprogesterone acetate or progesterone.
OBJECTIVE
In vivo and in vitro rat models of hormone therapy were used to test the following hypotheses: (1) estrogen acts directly on cerebrovascular estrogen receptors to increase endothelial nitric oxide synthase (eNOS); (2) increased protein correlates with higher NOS activity; and (3) effects of estrogen on eNOS are altered by concurrent treatment with either medroxyprogesterone acetate (MPA) or progesterone.


METHODS
Blood vessels were isolated from brains of ovariectomized female rats; some were treated for 1 month with estrogen, estrogen and progesterone, or estrogen and MPA. Isolated cerebral vessels were also treated in vitro with estrogen in the absence and presence of progesterone, MPA, tamoxifen, and the estrogen receptor antagonist ICI 182 780. Levels of eNOS were measured by Western blot, and NOS activity was measured by [14C]arginine-[14C]citrulline conversion.


RESULTS
Chronic hormone treatment in vivo resulted in plasma levels of 17beta-estradiol, progesterone, and MPA in the range of values found in humans. Estrogen treatment resulted in higher levels of cerebrovascular NOS activity that paralleled increases in eNOS protein. In vitro estrogen treatment for 18 hours also resulted in a concentration-dependent increase in eNOS protein (EC50 approximately 300 pmol/L) that was completely prevented by estrogen receptor antagonists tamoxifen or ICI 182 780. However, cotreatment with progesterone or MPA, either in vivo or in vitro, did not alter the effect of estrogen on eNOS protein.


CONCLUSIONS
Estrogen receptor activation in cerebrovascular tissue results in increased eNOS activity and protein levels. The latter effect persists in the presence of either progesterone or MPA. Thus, increased NO production by eNOS may contribute to the neuroprotective effects of estrogen.
McNeill
Anne Marie
AM
Department of Pharmacology, College of Medicine, University of California at Irvine, CA 92697-4625, USA.
Zhang
Chunying
C
Stanczyk
Frank Z
FZ
Duckles
Sue P
SP
Krause
Diana N
DN
D000818:Animals; D001808:Blood Vessels / Q000187:drug effects / Q000378:metabolism*; D001835:Body Weight / Q000187:drug effects; D001921:Brain / Q000098:blood supply; D004305:Dose-Response Relationship, Drug; D004343:Drug Implants; D004789:Enzyme Activation / Q000187:drug effects; D004958:Estradiol / Q000031:analogs & derivatives* / Q000097:blood / Q000494:pharmacology; D015914:Estrogen Replacement Therapy; D004967:Estrogens / Q000494:pharmacology*; D005260:Female; D000077267:Fulvestrant; D066298:In Vitro Techniques; D017258:Medroxyprogesterone Acetate / Q000008:administration & dosage / Q000097:blood / Q000494:pharmacology*; D023421:Models, Animal; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III; D009929:Organ Size / Q000187:drug effects; D010052:Ovariectomy; D011374:Progesterone / Q000097:blood / Q000494:pharmacology*; D051381:Rats; D011916:Rats, Inbred F344; D011960:Receptors, Estrogen / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013629:Tamoxifen / Q000494:pharmacology; D014599:Uterus / Q000187:drug effects
D004343:Drug Implants; D004967:Estrogens; D011960:Receptors, Estrogen; D013629:Tamoxifen; D000077267:Fulvestrant; D011374:Progesterone; D004958:Estradiol; D017258:Medroxyprogesterone Acetate; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; C496316:Nos3 protein, rat