266-POS

Abstract

Objectives Preeclampsia (preE) is a pregnancy disorder characterized by the de novo development of hypertension and proteinuria with multiple pathophysiologic triggers and mechanisms. Approximately 20% of the diabetic pregnant women develop preE. The mechanisms contributing to this effect is not well characterized. In a recent study, we show that hyperglycemia impairs cytotrophoblast (CTB) function via stress signaling. In the present study, we assess the apoptotic signaling mechanisms of excess glucose-induced CTBs dysfunction. Methods Human CTBs (Sw. 71) were treated with 0, 100, 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). Some cells were treated with D-Mannitol to evaluate the osmotic effect. Thereafter, cell lysates were utilized to measure PPARγ expression and p38 MAPK phosphorylation and apoptotic and stress signaling proteins; Bcl-2-associated X protein (Bax), anti-apoptotic Bcl-2 and caspase-3 and 9 and pro-inflammatory cyclooxygenase-2 (Cox-2) expression were assayed by western blot. Statistical comparisons were performed using analysis of variance with Duncan’s post hoc test. Results Both the p38 MAPK phosphorylation and PPARγ expression were upregulated (p 100 mg/dL glucose compared to basal (100 mg/dL). The expression of Bax/Bcl-2, Cox-2, caspase-3 and 9 were up-regulated (p Conclusions Hyperglycemia induced the apoptotic signaling in CTBs by upregulating the Bax/Bcl2, and caspase-3 and 9 Cox-2 expression. The attenuation of hyperglycemia-induced upregulation of apoptotic and stress signaling proteins by SB203580 or rosiglitazone pretreatment suggests the involvement of apoptotic signaling mechanisms in CTBs dysfunction. Disclosures C. Cawyer: None. D. Leonard: None. N. Drever: None. R.R. Jones: None. S.R. Allen: None. M.R. Beeram: None. T.J. Kuehl: None. M.N. Uddin: None.