title: Decidualisation of human endometrial stromal cells is associated with increased expression and secretion of prorenin.
abstract: BACKGROUND
  In pregnancy, the decidualised endometrium expresses high levels of prorenin and other genes of the renin-angiotensin system (RAS) pathway. In this study we aimed to determined if the RAS was present in endometrial stromal cells and if decidualisation upregulated the expression of prorenin, the prorenin receptor ((P)RR) and associated RAS pathways. Immortalised human endometrial stromal cells (HESCs) can be stimulated to decidualise by combined treatment with medroxyprogesterone acetate (MPA), 17β-estradiol (E2) and cAMP (MPA-mix) or with 5-aza-2'-deoxycytidine (AZA), a global demethylating agent.
  
  
  METHODS
  HESCs were incubated for 10 days with one of the following treatments: vehicle, MPA-mix, a combination of medroxyprogesterone acetate (MPA) and estradiol-17β alone, or AZA. Messenger RNA abundance and protein levels of prorenin (REN), the (P)RR (ATP6AP2), angiotensinogen (AGT), angiotensin converting enzyme (ACE), angiotensin II type 1 receptor (AGTR1), vascular endothelial growth factor (VEGF), and plasminogen activator inhibitor-1 (PAI-1) were measured by real-time PCR and ELISA's, respectively. Promyelocytic zinc finger (PLZF) and phospho-inositol-3 kinase (PIK3R1) mRNA abundances were also measured.
  
  
  RESULTS
  HESCs expressed the prorenin receptor (ATP6AP2), REN, AGT, ACE and low levels of AGTR1. MPA-mix and AZA stimulated expression of REN. Prorenin protein secretion was increased in MPA-mix treated HESCs. E2 + MPA had no effect on any RAS genes. MPA-mix treatment was associated with increased VEGF (VEGFA) and PAI-1 (SERPINE1) mRNA and VEGF protein.
  
  
  CONCLUSIONS
  An endometrial prorenin receptor/renin angiotensin system is activated by decidualisation. Since (P)RR is abundant, the increase in prorenin secretion could have stimulated VEGF A and SERPINE1 expression via Ang II, as both ACE and AGTR1 are present, or by Ang II independent pathways. Activation of the RAS in human endometrium with decidualisation, through stimulation of VEGF expression and secretion, could be critical in establishing an adequate blood supply to the developing maternal placental vascular bed.
authors.affiliation: School of Biomedical Sciences and Pharmacy, University of Newcastle, Newcastle, Australia and Mothers and Babies Research Centre, Hunter Medical Research Institute, Level 3 East, 1 Kookaburra Circuit, New Lambton Heights, NSW, 2305, Australia.
authors.forename: Eugenie R
authors.initials: ER
authors.lastname: Lumbers
authors.affiliation: School of Biomedical Sciences and Pharmacy, University of Newcastle, Newcastle, Australia and Mothers and Babies Research Centre, Hunter Medical Research Institute, Level 3 East, 1 Kookaburra Circuit, New Lambton Heights, NSW, 2305, Australia.
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.affiliation: School of Biomedical Sciences and Pharmacy, University of Newcastle, Newcastle, Australia and Mothers and Babies Research Centre, Hunter Medical Research Institute, Level 3 East, 1 Kookaburra Circuit, New Lambton Heights, NSW, 2305, Australia.
authors.forename: Sarah J
authors.initials: SJ
authors.lastname: Delforce
authors.affiliation: School of Biomedical Sciences and Pharmacy, University of Newcastle, Newcastle, Australia and Mothers and Babies Research Centre, Hunter Medical Research Institute, Level 3 East, 1 Kookaburra Circuit, New Lambton Heights, NSW, 2305, Australia.
authors.forename: Celine
authors.initials: C
authors.lastname: Corbisier de Meaultsart
authors.affiliation: The Liggins Institute, University of Auckland, Auckland, New Zealand.|Present address: Reproductive Sciences, Department of Obstetrics and Gynecology, School of Medicine, University of California San Francisco, San Francisco, USA.
authors.forename: Philip C
authors.initials: PC
authors.lastname: Logan
authors.affiliation: University of Queensland Centre for Clinical Research, University of QLD, St Lucia, QLD, Australia.
authors.forename: Murray D
authors.initials: MD
authors.lastname: Mitchell
authors.affiliation: School of Biomedical Sciences and Pharmacy, University of Newcastle, Newcastle, Australia and Mothers and Babies Research Centre, Hunter Medical Research Institute, Level 3 East, 1 Kookaburra Circuit, New Lambton Heights, NSW, 2305, Australia. Kirsty.pringle@newcastle.edu.au.
authors.forename: Kirsty G
authors.initials: KG
authors.lastname: Pringle
mesh_terms: D000242:Cyclic AMP / Q000494:pharmacology; D003841:Deoxycytidine / Q000494:pharmacology; D004717:Endometrium / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D006801:Humans; D017258:Medroxyprogesterone Acetate / Q000494:pharmacology; D010929:Placentation / Q000187:drug effects / Q000502:physiology; D017395:Plasminogen Activator Inhibitor 1 / Q000378:metabolism; D011247:Pregnancy; D011956:Receptors, Cell Surface / Q000378:metabolism; D012083:Renin / Q000378:metabolism*; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D000093342:Prorenin Receptor
chemical_list: D017395:Plasminogen Activator Inhibitor 1; D011956:Receptors, Cell Surface; C051321:SERPINE1 protein, human; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D003841:Deoxycytidine; D004958:Estradiol; D017258:Medroxyprogesterone Acetate; D000242:Cyclic AMP; D012083:Renin; D000093342:Prorenin Receptor