Prednisolone Alters Endometrial Decidual Cells and Affects Decidual-Trophoblast Interactions

Abstract

Poor pregnancy outcomes such as recurrent pregnancy loss (RPL) and preeclampsia are associated with impaired decidualization and abnormal trophoblast invasion. Emerging evidence suggests that use of corticosteroids, including prednisolone affects fertility by altering uterine function and may be associated with preeclampsia incidence. In this study, using primary and gestational-age appropriate tissue, we aimed to define the effect of prednisolone on human endometrial stromal fibroblast (hESF) decidualization and determine whether hESF decidualization in the presence of prednisolone would alter hESF regulation of trophoblast function. We found that prednisolone treatment reduced hESF cytokine expression (IL6, IL11, IL18, LIF, and LIFR) but had no effect on hESF expression or secretion of the classic markers of decidualization [prolactin (PRL) and IGFBP1]. Using proteomics we determined that prednisolone altered decidualized hESF protein production, enriching hESF proteins associated with acetylation and mitrochondria. Conditioned media from hESF decidualized in the presence of prednisolone significantly enhanced trophoblast outgrowth and trophoblast mRNA expression of cell motility gene PLCG1 and reduced trophoblast production of PGF. Prednisolone treatment during the menstrual cycle and 1st trimester of pregnancy might alter decidual interactions with other cells, including invasive trophoblast.

Keywords: decidualization; prednisolone; preeclampsia; recurrent pregnancy loss; trophoblast.

FIGURE 1

Prednisolone suppressedpro-inflammatory cytokine production by human endometrial stromalfibroblast (hESF). Prednisolone treatment: (A) inhibitedhESF pro-inflammatory cytokine [interleukin (IL) 6, and IL18] gene expression, but had no effect on IL10 gene expression, paired t-test, n = 3-4/group; (B) had no effect on hESF glucocorticoid receptor (GR) gene expression, paired t-test, n = 4/group; (C) had no effect on classic decidualization makers prolactin (PRL) or insulin-like growth factor binding protein (IGFBP)1 expression; (D) had no effect on decidualization genes bone morphogenic protein (BMP)2, BMP7, homeobox A (HOXA)10 or inhibinβA (INHBA), but inhibited IL11, leukemia inhibitory factor (LIF) and LIF receptor (LIFR) gene expression, paired t-test, n = 4/group. Data presented as mean ± SEM; *P < 0.05.

FIGURE 2

Prednisolone had no effect on classic decidualization markergene expression or secretion by decidualized human endometrial stromal fibroblast (hESF). Prednisolone treatment: (A,B) had no effect on decidualized hESF secretion of PRL or IGFBP1 from hESF (A) cultured fresh or (B) cultured from frozen stocks, repeated measures ANOVA; n = 3-4/group; (C–F) had no effect on decidualized hESF production of (C) GR; (D) classic decidualization markers PRL or IGFBP1; (E) decidualization genes BMP2, BMP7, HOXA10, IL11, INHBA, LIF or LIFR; (F) preeclampsia-associated genes endoglin (ENG), vascular endothelial growth factor receptor (FLT1) or placental-like growth factor (PGF), paired t-test, n = 4/group; Data presented as mean ± SEM; *P < 0.05.

FIGURE 3

Analysis of differently regulated decidualized human endometrial stromal fibroblast (hESF) proteins following in vitro decidualization in the presence of prednisolone. (A) Principal components analysis. (B) Heat-map. (C) Volcano plot. (D) Enriched pathways. Individual numbers of proteins identified is indicated in white at the base of each bar. Case: prednisolone treated hESF; Control: vehicle control treated hESF.

FIGURE 4

Prednisolone altered human endometrial stromal fibroblast (hESF) regulation of trophoblast function. (A) Extravillous trophoblast (EVT) outgrowth was significantly enhanced by conditioned media (CM) from hESF decidualized in the presence of prednisolone. Image shows representative outgrowth from villous tip. Insert shows area of outgrowth highlighted by dotted line. Paired t-test, n = 6. (B) EVT PLCG1 expression was significantly increased by treatment with CM from hESF decidualized in the presence of prednisolone. Paired t-test, n = 3/group. (C) EVT IL6 and IL18 expression was not altered by treatment with CM from hESF decidualized in the presence of prednisolone. Paired t-test, n = 3/group. (D) EVT PGF expression was significantly inhibited by treatment with CM from hESF decidualized in the presence of prednisolone. Paired t-test, n = 3/group. Data presented as mean ± SEM; *P < 0.05.