title: Attenuation of hyperglycemia-induced apoptotic signaling and anti-angiogenic milieu in cultured cytotrophoblast cells.
abstract: OBJECTIVE
  Preeclampsia (preE) is a hypertensive disorder that occurs 20% in diabetic pregnancy. We have shown that hyperglycemia impairs cytotrophoblast cell (CTB) function. In this study, we assess apoptotic and anti-angiogenic signaling in excess glucose-induced CTBs.
  
  
  METHODS
  Human extravillous CTBs (Sw. 71) were treated with 100, 150, 200, 300, or 400 mg/dL glucose for 48 h. Some cells were pretreated with a p38 inhibitor (SB203580) or a peroxisome proliferator-activated receptor gamma (PPARγ) ligand (rosiglitazone) or with D-mannitol. Cell lysates were utilized to measure p38 MAPK phosphorylation, PPARγ, Bcl-2-associated-X protein (Bax), anti-apoptotic Bcl-2, caspase-9, and cyclooxygenase-2 (Cox-2) expression by western blot. Levels of the vascular endothelial growth factor (VEGF), placental growth factor (PlGF), soluble fms-like tyrosine kinase-1 (sFlt-1), soluble endoglin (sEng), and interleukin 6 (IL-6) were measured in culture media using ELISA kits. Statistical comparisons were performed using analysis of variance with Duncan's post hoc test.
  
  
  RESULTS
  p38 phosphorylation and PPARγ were upregulated (p < 0.05) in CTBs treated with ≥150 mg/dL glucose compared to basal (100 mg/dL). Expressions of Bax/Bcl-2, Cox-2, and caspase-9 were upregulated (p < 0.05) in CTBs treated with ≥150 mg/dL glucose. Secretion of sFlt-1, sEng, and IL-6 was increased while VEGF and PIGF were decreased in CTB-treated ≥150 mg/dl of glucose (*p < 0.01 for each). SB203580 or rosiglitazone pretreatment significantly attenuated hyperglycemia-induced apoptotic and anti-angiogenic signaling. D-Mannitol had no effect.
  
  
  CONCLUSIONS
  Hyperglycemia induced apoptotic and anti-angiogenic signaling in CTBs. The observed diminution of hyperglycemia-induced signaling by SB203580 or rosiglitazone pretreatment suggests the involvement of apoptotic and anti-angiogenic signaling in CTB dysfunction.
authors.affiliation: a Departments of Obstetrics & Gynecology , Texas A&M Health Science Center College of Medicine/Baylor Scott & White Health , Temple , Texas , USA.
authors.forename: Chase
authors.initials: C
authors.lastname: Cawyer
authors.affiliation: b Medical Physiology , Texas A&M Health Science Center College of Medicine/Baylor Scott & White Health , Temple , Texas , USA.
authors.forename: Syeda H
authors.initials: SH
authors.lastname: Afroze
authors.affiliation: a Departments of Obstetrics & Gynecology , Texas A&M Health Science Center College of Medicine/Baylor Scott & White Health , Temple , Texas , USA.
authors.forename: Nathan
authors.initials: N
authors.lastname: Drever
authors.affiliation: a Departments of Obstetrics & Gynecology , Texas A&M Health Science Center College of Medicine/Baylor Scott & White Health , Temple , Texas , USA.
authors.forename: Steven
authors.initials: S
authors.lastname: Allen
authors.affiliation: a Departments of Obstetrics & Gynecology , Texas A&M Health Science Center College of Medicine/Baylor Scott & White Health , Temple , Texas , USA.
authors.forename: Richard
authors.initials: R
authors.lastname: Jones
authors.affiliation: b Medical Physiology , Texas A&M Health Science Center College of Medicine/Baylor Scott & White Health , Temple , Texas , USA.
authors.forename: David C
authors.initials: DC
authors.lastname: Zawieja
authors.affiliation: a Departments of Obstetrics & Gynecology , Texas A&M Health Science Center College of Medicine/Baylor Scott & White Health , Temple , Texas , USA.|c Pediatrics , Texas A&M Health Science Center College of Medicine/Baylor Scott & White Health , Temple , Texas , USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Kuehl
authors.affiliation: a Departments of Obstetrics & Gynecology , Texas A&M Health Science Center College of Medicine/Baylor Scott & White Health , Temple , Texas , USA.|c Pediatrics , Texas A&M Health Science Center College of Medicine/Baylor Scott & White Health , Temple , Texas , USA.|d Internal Medicine , Texas A&M Health Science Center College of Medicine/Baylor Scott & White Health , Temple , Texas , USA.
authors.forename: M Nasir
authors.initials: MN
authors.lastname: Uddin
mesh_terms: D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D053453:Caspase 9 / Q000378:metabolism; D002478:Cells, Cultured; D051546:Cyclooxygenase 2 / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; 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 / Q000378:metabolism; D000072483:Placenta Growth Factor / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D000077154:Rosiglitazone; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D045162:Thiazolidinediones / Q000494:pharmacology; D014327:Trophoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: C000595707:BCL2 protein, human; D004791:Enzyme Inhibitors; D007004:Hypoglycemic Agents; C508600:IL6 protein, human; D007093:Imidazoles; D015850:Interleukin-6; D047495:PPAR gamma; D019253:Proto-Oncogene Proteins c-bcl-2; D011725:Pyridines; D045162:Thiazolidinediones; D042461:Vascular Endothelial Growth Factor A; D051028:bcl-2-Associated X Protein; D000077154:Rosiglitazone; D000072483:Placenta Growth Factor; D051546:Cyclooxygenase 2; D040281:Vascular Endothelial Growth Factor Receptor-1; D048051:p38 Mitogen-Activated Protein Kinases; D053453:Caspase 9; D005947:Glucose; C093642:SB 203580
keywords: Anti-angiogenic; apoptotic; cytotrophoblast cells; diabetes; preeclampsia; pregnancy