Down-regulation of Rac-1 GTPase by Estrogen.
Rac1 GTPase is essential for the activation of the NAD(P)H oxidase complex and, thereby, regulates the release of reactive oxygen species (ROS) in the vessel wall. 17 beta-estradiol (E2) inhibits vascular ROS production. To elucidate the underlying molecular mechanisms we investigated the potential regulation of Rac1 by E2 in vascular smooth muscle cells. Treatment of vascular smooth muscle cells with angiotensin II as well as overexpression of the constitutively active mutant RacL61 increased ROS release as assessed by dichlorofluorescein fluorescence, whereas inhibition of Rac1 by Clostridium sordellii lethal toxin or overexpression of dominant-negative RacN17 inhibited ROS production. Treatment with E2 (100 nm) completely prevented angiotensin II-induced NAD(P)H oxidase activity and ROS production. E2 time and concentration dependently decreased angiotensin II-induced and basal Rac1 mRNA and protein expression as well as Rac1 activity. Down-regulation of Rac1 expression by E2 was mediated by inhibition of gene transcription (nuclear run-on assays), but E2 had no effect on Rac1 mRNA stability. Regulation of Rac1 was mediated by estrogen receptors since co-incubation with ICI 182.780 prevented down-regulation of Rac1. To test these observations in vivo, ovariectomized spontaneously hypertensive rats were treated with E2 or vehicle. Real-time PCR and Western blotting showed reduction of aortic Rac1 mRNA and protein by 32 and 58%, respectively. Furthermore, down-regulation of Rac1 by E2 was observed in human mononuclear cells of women with elevated E2 levels after controlled ovarian hyperstimulation. Rac1 GTPase gene-transcription and activity is regulated by 17 beta-estradiol, which may be an important molecular mechanism contributing to the cardiovascular effects of estrogens.
Laufs
Ulrich
U
Medizinische Klinik und Poliklinik, Innere Medizin III, Universitätskliniken des Saarlandes, 66421 Homburg/Saar, Germany. ulrich@laufs.com
Adam
Oliver
O
Strehlow
Kerstin
K
Wassmann
Sven
S
Konkol
Christian
C
Laufs
Kerstin
K
Schmidt
Werner
W
Böhm
Michael
M
Nickenig
Georg
G
D000804:Angiotensin II / Q000494:pharmacology*; D000818:Animals; D001013:Aorta, Thoracic / Q000201:enzymology; D002478:Cells, Cultured; D004958:Estradiol / Q000097:blood / Q000494:pharmacology*; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000201:enzymology / Q000378:metabolism; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000201:enzymology*; D010062:Ovulation Induction; D051381:Rats; D011918:Rats, Inbred SHR; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020830:rac1 GTP-Binding Protein / Q000097:blood / Q000235:genetics*
D017382:Reactive Oxygen Species; D000804:Angiotensin II; D004958:Estradiol; D020830:rac1 GTP-Binding Protein