Research
Obstetrics
Hyperglycemia impairs cytotrophoblast function via stress signaling

Presented in oral format at the 34th annual meeting of the Society for Maternal-Fetal Medicine, New Orleans, LA, Feb. 3-8, 2014.
https://doi.org/10.1016/j.ajog.2014.04.033Get rights and content

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.

Study Design

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.

Conclusion

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.

Section snippets

CTB cell culture

The human extravillous CTB cell line Sw.71 utilized in these studies was derived from first-trimester chorionic villus tissue and was kindly provided by Dr Gil G. Mor (Yale University School of Medicine, New Haven, CT). These cells are well characterized and share many characteristics with isolated primary cells, including the expression of cytokeratin-7, human leukocyte antigen class I antigen, human leukocyte antigen-G, BC-1, CD9, human chorionic gonadotropin, and human placental lactogen.

Hyperglycemia up-regulated p38 MAPK phosphorylation and PPAR-γ expression

Figure 2 demonstrates that the ratio of phosphorylated p38 MAPK to the nonphosphorylated p38α/β was significantly (P < .05) up-regulated in all the CTB cell cultures treated with 495 mg/dL or more of glucose compared with basal (45 mg/dL). Intermediate levels of glucose exposure, beginning at more than 225 mg/dL, produced intermediate changes that were not different from values at either exposure to 45 mg/dL or 495 mg/dL. In addition, Figure 2 shows that PPAR-γ expression was significantly

Comment

As confirmed by multiple other investigators, preeclampsia is commonly associated with inadequate endovascular cytotrophoblast function.8, 9 Postulating supraphysiological hyperglycemia as an environmental stressor, we demonstrate an induction of PPAR-γ expression that is consistent with that reported by Suwaki et al42 yet differing with Jawerbaum et al.43 These results also are consistent with those reported by Zhou et al,40 who illustrated that hyperglycemia activates multiple MAPK pathways

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    This study was supported by the Scott, Sherwood, and Brindley Foundation, Department of Obstetrics and Gynecology (M.N.U.), and the Noble Centennial Endowment for Research in Obstetrics and Gynecology (T.J.K.), Scott and White Healthcare, Temple, TX.

    The authors report no conflict of interest.

    Cite this article as: Cawyer CR, Horvat D, Leonard D, et al. Hyperglycemia impairs cytotrophoblast function via stress signaling. Am J Obstet Gynecol 2014;211:541.e1-8.

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