title: Estrogen attenuates integrin-beta(3)-dependent adventitial fibroblast migration after inhibition of osteopontin production in vascular smooth muscle cells.
abstract: BACKGROUND
  Previous in vitro studies have suggested that estrogen attenuates the vascular injury response by modulating vascular smooth muscle cell (VSMC) expression of soluble factor(s) directing migration of adventitial fibroblasts. Previous in vivo studies have established a role for osteopontin (OPN) and its integrin receptors after vascular injury. In this study, we examined OPN expression in activated VSMCs, its modulation by estrogen, and its effects on adventitial fibroblast migration. In addition, the relative functional roles of beta(1)- and beta(3)-integrin-matrix interactions were examined.
  
  
  RESULTS
  Primary cultures of VSMCs and adventitial fibroblasts were derived from female Sprague-Dawley rats. Serum-activated VSMCs expressed high levels of OPN mRNA and secreted protein that was effectively inhibited by estrogen treatment (10(-7) mol/L). Compared with VSMCs, fibroblasts expressed similar levels of integrins alphanu and beta(1) and higher levels of integrin-beta(3). Exogenous OPN (5.0 to 40 microg/mL) directed fibroblast migration in a dose-dependent fashion. Anti-beta(3)-integrin antibody (F11) pretreatment markedly inhibited adventitial fibroblast migration directed by exogenous OPN or VSMC-conditioned medium in a dose-dependent manner. In contrast, anti-beta(1)-integrin antibody (Ha2/5) did not affect fibroblast migration. Similarly, pretreatment with either linear or cyclic RGD peptides (10 to 1000 micromol/L) inhibited fibroblast migration directed by OPN or VSMC-conditioned medium in a dose-dependent manner.
  
  
  CONCLUSIONS
  These observations suggest that estrogen indirectly attenuates integrin-beta(3)-dependent adventitial fibroblast migration after inhibition of OPN expression in VSMCs.
authors.affiliation: Departments of Medicine, Vascular Biology and Hypertension Program, and Surgery, Division of Transplantation, University of Alabama at Birmingham, Birmingham, Alabama, USA.
authors.forename: G
authors.initials: G
authors.lastname: Li
authors.forename: Y F
authors.initials: YF
authors.lastname: Chen
authors.forename: S S
authors.initials: SS
authors.lastname: Kelpke
authors.forename: S
authors.initials: S
authors.lastname: Oparil
authors.forename: J A
authors.initials: JA
authors.lastname: Thompson
mesh_terms: D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D015703:Antigens, CD / Q000276:immunology / Q000502:physiology*; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004958:Estradiol / Q000494:pharmacology; D004967:Estrogens / Q000502:physiology*; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000502:physiology*; D039661:Integrin beta3; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D009842:Oligopeptides / Q000494:pharmacology; D053495:Osteopontin; D010980:Platelet Membrane Glycoproteins / Q000276:immunology / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012795:Sialoglycoproteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000494:pharmacology
chemical_list: D000906:Antibodies; D015703:Antigens, CD; D004967:Estrogens; D039661:Integrin beta3; D009842:Oligopeptides; D010980:Platelet Membrane Glycoproteins; D012333:RNA, Messenger; D012795:Sialoglycoproteins; C506897:Spp1 protein, rat; D053495:Osteopontin; D004958:Estradiol; C047981:arginyl-glycyl-aspartic acid