title: Hyperglycemia impairs cytotrophoblast function via stress signaling.
abstract: OBJECTIVE
  Diabetes mellitus is a risk factor for preeclampsia. Cytotrophoblast (CTB) invasion is facilitated from the conversion of plasminogen to plasmin by urokinase plasminogen activator (uPA), regulated by plasminogen activator inhibitor 1 (PAI-1), and may be inhibited in preeclampsia. This study assessed signaling mechanisms of hyperglycemia-induced CTB dysfunction.
  
  
  METHODS
  Human CTBs were treated with 45, 135, 225, 495, or 945 mg/dL glucose for 48 hours. Some cells were pretreated with a p38 inhibitor (SB203580) or a peroxisome proliferator-activated receptor-gamma (PPAR-γ) ligand (rosiglitazone). Expression of uPA, PAI-1, and PPAR-γ levels and p38 mitogen-activated protein kinase phosphorylation were measured by Western blot in cell lysates. Messenger ribonucleic acid of uPA and PAI-1 was measured by quantitative polymerase chain reaction. Levels of interleukin-6, angiogenic (vascular endothelial growth factor [VEGF], placenta growth factor [PlGF]) and antiangiogenic factors (soluble fms-like tyrosine kinase-1 [sFlt-1], soluble endoglin [sEng]) were measured in the media by enzyme-linked immunosorbent assay kits. Statistical comparisons were performed using analysis of variance with a Duncan's post-hoc test.
  
  
  RESULTS
  Both uPA and PAI-1 protein and messenger ribonucleic acid were down-regulated (P < .05) in CTBs treated with 135 mg/dL glucose or greater compared with basal (45 mg/dL). The sEng, sFlt-1, and interleukin-6 were up-regulated, whereas the VEGF and PlGF were down-regulated by 135 mg/dL glucose or greater. p38 phosphorylation and PPAR-γ were up-regulated (P < .05) in hyperglycemia-treated CTBs. The SB203580 or rosiglitazone pretreatment showed an attenuation of glucose-induced down-regulation of uPA and PAI-1.
  
  
  CONCLUSIONS
  Hyperglycemia disrupts the invasive profile of CTB by decreasing uPA and PAI-1 expression; down-regulating VEGF and PlGF; and up-regulating sEng, sFlt-1, and interleukin-6. Attenuation of CTB dysfunction by SB203580 or rosiglitazone pretreatment suggests the involvement of stress signaling.
authors.affiliation: Department of Obstetrics and Gynecology, Scott and White Healthcare/Texas A&M Health Science Center College of Medicine, Temple, TX.
authors.forename: Chase R
authors.initials: CR
authors.lastname: Cawyer
authors.affiliation: Department of Obstetrics and Gynecology, Scott and White Healthcare/Texas A&M Health Science Center College of Medicine, Temple, TX.
authors.forename: Darijana
authors.initials: D
authors.lastname: Horvat
authors.affiliation: Prehealth Studies, Baylor University, Waco, TX.
authors.forename: Dean
authors.initials: D
authors.lastname: Leonard
authors.affiliation: Department of Obstetrics and Gynecology, Scott and White Healthcare/Texas A&M Health Science Center College of Medicine, Temple, TX.
authors.forename: Steven R
authors.initials: SR
authors.lastname: Allen
authors.affiliation: Department of Obstetrics and Gynecology, Scott and White Healthcare/Texas A&M Health Science Center College of Medicine, Temple, TX.
authors.forename: Richard O
authors.initials: RO
authors.lastname: Jones
authors.affiliation: Department of Medical Physiology, Scott and White Healthcare/Texas A&M Health Science Center College of Medicine, Temple, TX.
authors.forename: David C
authors.initials: DC
authors.lastname: Zawieja
authors.affiliation: Department of Obstetrics and Gynecology, Scott and White Healthcare/Texas A&M Health Science Center College of Medicine, Temple, TX.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Kuehl
authors.affiliation: Department of Obstetrics and Gynecology, Scott and White Healthcare/Texas A&M Health Science Center College of Medicine, Temple, TX. Electronic address: mnuddin@sw.org.
authors.forename: Mohammad N
authors.initials: MN
authors.lastname: Uddin
mesh_terms: D015703:Antigens, CD / Q000187:drug effects / Q000378:metabolism; D016640:Diabetes, Gestational / Q000378:metabolism; D000071063:Endoglin; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D006943:Hyperglycemia / Q000378:metabolism*; D007004:Hypoglycemic Agents / Q000494:pharmacology; D007093:Imidazoles / Q000494:pharmacology; D015850:Interleukin-6 / Q000378:metabolism; D047495:PPAR gamma / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D000072483:Placenta Growth Factor; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000187:drug effects / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism; D000077154:Rosiglitazone; D015398:Signal Transduction / Q000187:drug effects; D045162:Thiazolidinediones / Q000494:pharmacology; D014327:Trophoblasts / Q000187:drug effects*; D014568:Urokinase-Type Plasminogen Activator / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000187:drug effects / Q000378:metabolism; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000187:drug effects / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000187:drug effects / Q000378:metabolism
chemical_list: D015703:Antigens, CD; C064206:ENG protein, human; D000071063:Endoglin; D007004:Hypoglycemic Agents; C508600:IL6 protein, human; D007093:Imidazoles; D015850:Interleukin-6; C000608036:PGF protein, human; D047495:PPAR gamma; D017395:Plasminogen Activator Inhibitor 1; D011257:Pregnancy Proteins; D011725:Pyridines; D011956:Receptors, Cell Surface; C051321:SERPINE1 protein, human; D045162:Thiazolidinediones; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D000077154:Rosiglitazone; D000072483:Placenta Growth Factor; C501162:FLT1 protein, human; D040281:Vascular Endothelial Growth Factor Receptor-1; D048051:p38 Mitogen-Activated Protein Kinases; D014568:Urokinase-Type Plasminogen Activator; D005947:Glucose; C093642:SB 203580
keywords: angiogenesis; cell invasion; cytotrophoblast cells; hyperglycemia