title: Pregnancy-stimulated growth of vascular smooth muscle cells: importance of protein kinase C-dependent synergy between estrogen and platelet-derived growth factor.
abstract: Dramatic smooth muscle cell (SMC) growth occurs in the uterine artery during pregnancy. The potential for pregnancy-associated growth may also exist at other vascular sites. We tested the hypothesis that increased growth of uterine artery SMC isolated from pregnant (vs. nonpregnant) guinea pigs would be detectable in culture, that pregnancy-associated phenotypic changes would also be found in nonuterine vascular cells (aortic SMC), and that the enhanced growth would be dependent on estrogen, peptide growth factors like platelet-derived growth factor (PDGF), and protein kinase C (PKC). Growth responses were measured by [3H]-thymidine incorporation and cell counts. Uterine artery SMC from pregnant guinea pigs grew to a higher plateau density with serum stimulation, had increased spontaneous DNA synthesis and persistent growth following serum with-drawal, and were more responsive to 3-30 ng/ml PDGF-BB than nonpregnant cells. Aortic SMC from pregnant animals also grew to a higher plateau density and had enhanced responsiveness of PDGF-BB. This increased response to PDGF-BB by pregnant uterine artery and aortic SMC (40-233% increase over nonpregnant PDGF result) was reproduced in nonpregnant cells by pretreatment for 1-24 h with 17-beta(beta)-estradiol (30-100 nM). Neither the pregnancy-induced difference nor the estradiol pretreatment was associated with increased PDGF-BB binding activity. The synergistic effect of 17 beta-estradiol was partially (62%) reproduced with 17-alpha(alpha)-estradiol, an isomer which does not bind the estrogen receptor. This suggested that 17 beta-estradiol modulates the PDGF-BB response by both estrogen-receptor- and nonreceptor-mediated mechanisms. To test if the estrogen effects were dependent on PKC, two different antagonist strategies (3 microM dihydrosphingosine and phorbol-ester-induced downregulation) were applied prior to 17 alpha- or beta-estradiol and blocked the enhanced responses to PDGF. The synergistic effect of 17 beta-estradiol on PDGF was then reproduced by 1 h pretreatment with the cell-permeable PKC activator, 10 nM PMA. We conclude that pregnancy stimulates increased growth of uterine and aortic SMC in vitro which is dependent on estrogen, PDGF, and PKC and may be important in vascular remodeling during pregnancy.
authors.affiliation: Cardiovascular Pulmonary Research Laboratory, University of Colorado Health Sciences Center, Denver 80262, USA.
authors.forename: L E
authors.initials: LE
authors.lastname: Keyes
authors.forename: L G
authors.initials: LG
authors.lastname: Moore
authors.forename: S J
authors.initials: SJ
authors.lastname: Walchak
authors.forename: E C
authors.initials: EC
authors.lastname: Dempsey
mesh_terms: D000818:Animals; D001011:Aorta / Q000166:cytology; D001798:Blood Proteins / Q000494:pharmacology; D002455:Cell Division / Q000502:physiology; D004247:DNA / Q000096:biosynthesis; D004357:Drug Synergism; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D006168:Guinea Pigs; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000201:enzymology / Q000648:ultrastructure; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology*; D011247:Pregnancy; D011485:Protein Binding / Q000502:physiology; D011493:Protein Kinase C / Q000378:metabolism*; D011960:Receptors, Estrogen / Q000378:metabolism; D017479:Receptors, Platelet-Derived Growth Factor / Q000378:metabolism; D013936:Thymidine / Q000378:metabolism; D014316:Tritium / Q000378:metabolism; D014599:Uterus / Q000098:blood supply / Q000166:cytology
chemical_list: D001798:Blood Proteins; D010982:Platelet-Derived Growth Factor; D011960:Receptors, Estrogen; D014316:Tritium; D004958:Estradiol; D004247:DNA; D017479:Receptors, Platelet-Derived Growth Factor; D011493:Protein Kinase C; D013936:Thymidine