This Article Abstract Full Text (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Strakova, Z. Articles by Fazleabas, A. T. Search for Related Content PubMed PubMed Citation Articles by Strakova, Z. Articles by Fazleabas, A. T.

Interleukin-1ß Induces the Expression of Insulin-Like Growth Factor Binding Protein-1 during Decidualization in the Primate¹

Department of Obstetrics and Gynecology, University of Illinois at Chicago, Chicago, Illinois 60212-7313

Address all correspondence and requests for reprints to: Asgi T. Fazleabas, Ph.D., The University of Illinois at Chicago, Department of Obstetrics and Gynecology, 820 South Wood Street (M/C 808), Chicago, Illinois 60612-7313. E-mail: asgi{at}uic.edu

	Abstract

Top Abstract Introduction Materials and Methods Results Discussion References

 
Since interleukin (IL)-1 can modulate fetal/maternal interactions, we hypothesized that IL-1ß is one potential embryonic cytokine that regulates the conceptus-induced decidual response in baboon stromal fibroblasts. Treatment of stromal fibroblasts with IL-1ß (10 ng/ml, 10 min) resulted in the phosphorylation of p38 mitogen-activated protein kinase and IB-. This suggests that IL-1ß induces multiple signaling pathways in stromal cells that result in the activation of mitogen-activated protein kinase cascade and the transcription factor NF-B. After 4 h of stimulation, IL-1ß induced gene expression of cyclooxygenase-2 (COX-2) but not cyclooxygenase-1 (COX-1). PGE₂ synthesis paralleled COX-2 messenger RNA expression. The addition of hormones [36 nM estradiol-17ß, 1 µM medroxyprogesterone acetate, and 100ng/ml relaxin] to IL-1ß-treated cells induced insulin-like growth factor binding protein-1 (IGFBP-1) messenger RNA expression after 3 days of incubation. A specific COX-2 inhibitor, NS 398 (10 nM), partially inhibited IGFBP-1 protein synthesis. In contrast, the induction of IGFBP-1 by N⁶, 2'-O-dibutyryladenosine 3:5'-cyclic monophosphate (dbcAMP) and hormones was not affected by NS 398 treatment. Both dbcAMP and IL-1ß, in the presence of hormones, can independently induce IGFBP-1 gene expression and decidualization. However, if IL-1ß and dbcAMP were added together, IGFBP-1 expression was inhibited. These data suggest that IL-1ß can activate multiple signaling pathways that either positively or negatively regulate IGFBP-1 gene expression and decidualization.

	Introduction

Top Abstract Introduction Materials and Methods Results Discussion References

 
IN PRIMATES AND rodents, the uterine endometrial stromal cells differentiate to decidual cells after the establishment of pregnancy. Decidual cells play an important role in implantation and provide nutritional support for embryo. Decidual cells are also thought to produce factors that control trophoblast invasion and protect the embryo from maternal immune rejection. During the process of decidualization in the primate, fibroblast-like stromal cells change morphologically into polygonal cells and begin to express specific decidual proteins (1, 2). The major secretory product of the human and baboon decidualized endometrium is insulin-like growth factor binding protein-1 (IGFBP-1; 3, 4, 5).

Using a simulated pregnant baboon model, we demonstrated that exogenous CG, in conjunction with estrogen and progesterone, can initiate functional changes in uterine endometrium that are comparable with those seen during early pregnancy (2, 6). However, the hormone treatments alone were not sufficient to complete the decidualization process, suggesting the involvement of a conceptus factor (7, 8).

Implantation has been characterized as an inflammatory type response. A number of cytokines have been identified at the implantation site. Interleukin (IL)-1 was identified as one such paracrine factor that modulates the communication between the maternal endometrium and embryo (9, 10).

The cyclooxygenase (COX) enzymes required for PG biosynthesis exist in two isoforms (constitutive COX-1 and inducible COX-2). Studies using COX-2 null mice show reproductive failure at ovulation, fertilization, implantation, and decidualization (11). In the baboon endometrium, with the onset of pregnancy, COX-2 is expressed specifically in the stromal cells at the site of implantation (12). IL-1 regulates COX-2 gene expression in a variety of cell types, including stromal fibroblasts (13, 14).

Therefore, the purpose of our study was to evaluate the role of IL-1 as a component of the decidualization process. Specifically we chose to study its ability to induce signal transduction and regulate COX-2 and IGFBP-1 expression in stromal fibroblasts isolated from baboons. Comparative studies were also done with human stromal fibroblasts (HuF).

	Materials and Methods

Top Abstract Introduction Materials and Methods Results Discussion References

 
Materials
Recombinant human IL-1ß and IL-1 receptor antagonist (IL-1ra) were obtained from R&D Systems, Inc. (Minneapolis, MN). Rabbit polyclonal phospho-p38 mitogen-activated protein kinase (MAPK) (Thr¹⁸⁰/Tyr¹⁸²), p38 MAPK, phospho-IB- (Ser³²), IB- antibodies were from New England Biolabs, Inc. (Beverly, MA). The selective inhibitor of COX-2, NS-398 [N-(2-Cyclohexyloxy-4-nitrophenyl) methanesulfonamide] was from Calbiochem (San Diego, CA); N⁶, 2'-O-dibutyryladenosine 3:5'-cyclic monophosphate (dbcAMP) was from Sigma (St. Louis, MO). All cell culture supplies were obtained from Life Technologies, Inc. (Gaithersburg, MD). Other reagents of cell culture grade were purchased from Fisher Scientific (Itasca, IL), Sigma (St. Louis, MO), or Roche Molecular Biochemicals (Indianapolis, IN).

Isolation of baboon stromal cells
Midluteal phase (9–12 days post ovulation) endometrial tissue was obtained from adult female baboons (Papio anubis) by endometrectomy or after hysterectomy. All animal studies were approved by the Animal Care Committee at the University of Illinois at Chicago. Stromal cells were isolated from baboon endometrial tissues as described in detail previously (8).

Isolation of human fibroblast cells
Decidualized uterine endometrium maintains a proliferating population of fibroblastic cells, which closely resemble the stromal cells (15, 16). HuF were isolated from decidua parietalis dissected from the placental membranes after normal vaginal delivery at term. These studies were approved by the Institutional Review Board of the University of Illinois. Briefly, scraped cells were digested in 0.1% collagenase, 0.02% deoxynuclease in calcium- and magnesium-free HBSS. Cells were isolated in the same manner as described previously for baboon stromal fibroblasts (8). Cells were plated in four 100-mm culture dishes (Becton Dickinson and Co. Labware, Franklin Lakes, NJ) and placed into an incubator at 37 C, 5% CO₂. The next day, the plates were extensively washed with PBS to remove nonadherent (mainly decidual) cells. At confluence, cells were trypsinized and used for experiments in passage number 3–5. Cell purity was assessed by immunocytochemistry using antibodies against cytokeratin (DAKO Corp., Carpenteria, CA), vimentin (Zymed Laboratories, Inc., San Francisco, CA). The purity of the fibroblast cell preparations used in studies was more than 95%.

Treatment of cells
Experiments were performed when the baboon stromal cells or HuF cells reached 80–90% confluence. Culture medium was changed into serum-free culture medium for the 1-, 4-, and 24-h time points or into 2% stripped FBS culture medium for longer incubation times. The drug treatments are indicated in figure legends. Cell culture medium was changed every 2 days. Twenty-four hours before final experimental end point, the medium was changed into serum-free culture medium. The word "hormones" in this paper includes treatment with a mix of (final concentration) 36 nM estradiol-17ß, 1 µM medroxyprogesterone acetate, and 100 ng/ml highly purified porcine relaxin.This concentration of relaxin does not significantly increase intracellular levels of cAMP after 15 min of incubation (control, 0.79 ± 0.39 pmol/µg protein; relaxin, 1.43 ± 0.50 pmol/µg protein). In the experiments with NS 398, the cells were pretreated with inhibitor for 1 h and then exposed to other treatments. At each time point, the medium was collected, and the IGFBP-1 present in the culture medium was measured using an enzyme-linked immunosorbent assay (ELISA) kit (Diagnostic Systems Laboratories, Inc. Webster, TX). Cells were lysed with TriReagent (Molecular Research Center, Inc., Cincinnati, OH), and RNA was extracted using the protocol provided by the manufacturer. Cellular protein content was measured using the Micro BCA protein assay kit (Pierce Chemical Co., Rockford, IL).

PGE₂ assay
The PGE₂ concentration in cell culture medium was estimated using a PGE₂ enzyme immunoassay kit from Amersham Pharmacia Biotech (Arlington Heights, IL). The sensitivity of the assay was 2.5 pg/ml of medium.

Preparation of cell lysates and immunodetection
Stromal cells were grown to confluence on 35-mm (diameter) dishes and maintained in serum-free medium for 18 h. After each of the respective treatments, the cells were rinsed twice with ice-cold PBS, pH 7.4, and lysed on ice with 150 µl lysis buffer, as previously described (17). Cell lysate proteins (100 µg) were subjected to 10% SDS-PAGE and were transferred into polyvinylidene difluoride (PVDF) membranes. The membranes were incubated in PBS containing 3% BSA and 0.1% Tween 20 for 1 h, followed by overnight incubation with primary antibody (see legends in Fig. 1 and Fig. 8, A and B, for specific antibodies). The membranes were rinsed and incubated with the secondary IgG antibody labeled with horseradish peroxidase for 1 h. Immunocomplexes were visualized by enhanced chemiluminescence (Amersham Pharmacia Biotech).

View larger version (46K): [in this window] [in a new window]   Figure 1. IL-1ß activates p38 MAPK and IB- phosphorylation in baboon stromal cells A, Confluent baboon stromal cells were treated with IL-1ß (10 ng/ml) for 5, 10, and 20 min or with IL-1ra (10 ng/ml) for 10 min. Proteins from cell lysates were separated on 10% SDS-PAGE gels and transferred to PVDF membranes. Western blots were probed with a phospho-p38 MAPK antibody (top panel) and membrane was reprobed with p38 MAPK antibody (bottom panel). Note that p38 was only phosphorylated by IL-1ß and not by IL-1 receptor antagonist. B, Confluent baboon stromal cells were treated with IL-1ß (10 ng/ml) and IL-1ra (10 ng/ml) for the indicated times. The IB- protein was detected in cell lysates by Western blots using a phospho-IB- (Ser 32) antibody (top panel) and reprobed with IB- antibody (bottom panel). Note that IB- protein was phosphorylated only by IL-1ß and not by IL-1 receptor antagonist.

View larger version (26K): [in this window] [in a new window]   Figure 8. Signal transduction and IGFBP-1 synthesis induced by IL-1ß in human fibroblasts. A, Confluent human fibroblast cells were treated with IL-1ß (10 ng/ml) for 10 and 20 min or with IL-1ra (10 ng/ml) for 10 min. Proteins from cell lysates were separated on 10% SDS-PAGE gels and transferred to PVDF membranes. Western blots were probed with a phospho-p38 MAPK antibody (top panel), and membrane was reprobed with p38 MAPK antibody (bottom panel). Note that p38 was only phosphorylated by IL-1ß and not by IL-1 receptor antagonist. B, Confluent human fibroblast cells were treated with IL-1ß (10 ng/ml) and IL-1ra (10 ng/ml) for the indicated times. The IB- protein was detected in cell lysates by Western blots using a phospho-IB- (Ser 32) antibody (top panel) and reprobed with IB- antibody (bottom panel). Note that IB- protein was phosphorylated only by IL-1ß and not by IL-1 receptor antagonist. Compare data in A and B with data shown in Fig. 1C. Confluent human fibroblast cells were treated with IL-1ß (10 ng/ml), hormones, and 0.1 mM dbcAMP in different combinations, as indicated in the figure for 13 days, as described in Materials and Methods. Cell supernatants from the last day of the experiment were analyzed for IGFBP-1 protein, by ELISA. The values are mean ± SEM of three separate experiments done in triplicate. Significant differences, at P < 0.05, between groups, are designated by different lowercase letters. Note the decrease in IGFBP-1 secretion after the addition of IL-1ß to the dbcAMP and hormone-treated cells, which was comparable with that seen in baboon stromal cells (Fig. 7).

Statistical analyses
One-way ANOVA was used to test the null hypothesis of group differences, followed by a two-tailed Student’s t test for pairwise comparisons. Each experiment was repeated three times in triplicate, and a P value of <0.05 was considered significant.

	Results

Top Abstract Introduction Materials and Methods Results Discussion References

 
IL-1ß activates immediate signal response in baboon stromal cells
MAPKs are one of the major signaling systems by which cells transduce extracellular signals into intracellular responses. IL-1ß activates p38 kinase subfamily of MAPK in baboon stromal cells. In cell lysates, the presence of the phosphorylated form of p38 kinase was detected after 10 and 20 min of IL-1ß (10 ng/ml) stimulation. The phosphorylated form of p38 MAPK was not detected after 10 min of stimulation with IL-1ra (10 ng/ml; Fig. 1A).

In most mammalian cells, IB- serves as a prevalent inhibitor of NF-B complex (18). Activation of IB-, via phosphorylation at the Ser³² and Ser³⁶ residues, results in the release and nuclear translocation of active NF-B (19). The immunodetection with phospho-IB- (Ser³²) antibody revealed the presence of activated IB- protein after 5 and 10 min of IL-1ß treatment but not IL-1ra treatment (Fig. 1B). These data suggest that both p38 MAPK and IB- are phosphorylated and activated by IL-1ß in baboon stromal cells.

IL-1ß induces COX-2, but not COX-1 gene expression, in stromal cells
The expression of COX-1 and COX-2 messenger RNA (mRNA), after 1, 4, and 24 h of IL-1ß (10 ng/ml) treatment, was determined by RT-PCR in baboon stromal fibroblasts (Fig 2). COX-1 mRNA was evident in cells with no treatment, and no further regulation by IL-1ß was detected during culture (Fig. 2A). In contrast, after 4 h of IL-1ß stimulation, there was an increase in COX-2 mRNA, which decreased by 24 h of incubation (Fig. 2B). Treatment with IL-1 antagonist (IL-1ra, 10 ng/ml), which binds to the IL-1 receptor but does not activate the signaling pathways, did not induce COX-2 expression. As a positive control for COX-2 expression, human endometrial epithelial cells (HES) were treated for 4 h with IL-1ß (13).

View larger version (34K): [in this window] [in a new window]   Figure 2. IL-1ß induces COX-2 gene expression. Baboon stromal cells were cultured to 90% confluence and treated in serum-free medium with IL-1ß (10 ng/ml), IL-1 receptor antagonist (IL-1ra; 10 ng/ml), or without any additions for 1 h, 4 h, and 24 h, as indicated in the figure. RT-PCR, using primers for COX-1 and H3.3 (A) or COX-2 and H3.3 (B), was performed on isolated RNA. HES were used as positive control. The densitometric scans of the representative gels from two separate experiments are shown at the bottom of each of the gels. The data are expressed in arbitrary units as a ratio of COX-1/H3.3 or COX-2/H3.3. Note that COX-1 expression was not changed, but there is a transient increase in COX-2 expression after 4 h of IL-1ß treatment.

View larger version (12K): [in this window] [in a new window]   Figure 3. Production of PGE2 after IL-1ß stimulation. Stromal cells, isolated from cycling baboons, were treated in the absence or presence of IL-1ß (10 ng/ml), IL-1ra (10 ng/ml), and a combination of IL-1ß and IL-ra for 1, 4, and 24 h. Cell culture medium was assayed for PGE2 using an enzyme immunoassay (Biotrak EIA system, Amersham Pharmacia Biotech). Data are expressed as the mean ± SEM of three different experiments. Significant differences, at P < 0.05, between groups, are designated by different lowercase letters. Note the increase in PGE2 level after 24 h of IL-1ß treatment and partial inhibition of this increase in the presence of IL-ra.

Effect of IL-1ß on -SMA and IGFBP-1 expression

View larger version (36K): [in this window] [in a new window]   Figure 4. Effect of IL-1ß on -SMA and IGFBP-1 expression. Baboon stromal cells were treated for 6 h, 3 days, and 6 days with IL-1ß, IL-1ß plus hormones (36 nM 17ß-estradiol, 1 µM medroxyprogesterone acetate, and 100 ng/ml relaxin), hormones alone, or 0.1 mM dbcAMP and hormones. Total RNA was isolated and subjected to the RT-PCR using primers for -SMA, IGFBP-1, and the internal standard H3.3 in a single tube (A). B, Densitometric evaluation of gel expressed in arbitrary units of -SMA/H3.3 values and IGFBP-1/H3.3. Note the increase in IGFBP-1 expression after IL-1ß treatment in the presence of hormones, which was accompanied by a gradual decrease in -SMA expression, after 6 days of incubation.

View larger version (11K): [in this window] [in a new window]   Figure 5. IGFPB-1 secretion in response to decidual stimuli. Baboon stromal cells were grown to 90% confluence and treated with IL-1ß (10 ng/ml); IL-1ß in the presence of hormones, 0.1 mM dbcAMP and hormones, or hormones alone for 3 days and 6 days, as indicated in the figure. The IGFBP-1 protein released into the cell culture media was detected by a specific ELISA. The values are mean ± SEM of three separate experiments done in triplicate. Significant differences, at P < 0.05, between groups, are designated by different lowercase letters. Note the significant increase in IGFBP-1 levels after 6 days of IL-1ß treatment in the presence of hormones. The IGFBP-1 concentration induced by IL-1ß and hormones is significantly higher, in comparison with treatment with hormones only, and lower than dbcAMP treatment in the presence of hormones.

View larger version (10K): [in this window] [in a new window]   Figure 6. Effect of COX-2 inhibitor on IGFBP-1 secretion. Baboon stromal fibroblasts were cultured without, or were pretreated with, a COX-2 inhibitor, NS-398 (10 nM) for 1 h. This preincubation was followed by stimulation with IL-1ß (10 ng/ml) and hormones (A) or 0.1 mM dbcAMP and hormones (B) for 3 or 6 days. The cell culture media were analyzed by ELISA for IGFBP-1 protein levels. The values are mean ± SEM of three separate experiments done in triplicate. Significant differences, at P < 0.05, between groups, are designated by different lowercase letters. Note that pretreatment with NS-398 did not influence IGFBP-1 induced by dbcAMP and hormones but caused significant decrease in IL-1ß and hormone-induced IGFBP-1 protein.

View larger version (13K): [in this window] [in a new window]   Figure 7. IL-1ß inhibits dbcAMP and hormone-induced IGFBP-1 secretion. Confluent baboon stromal cells were treated with IL-1ß (10 ng/ml), hormones (H), and 0.1 mM dbcAMP in different combinations, as indicated in the figure, for 6 days, as described in Materials and Methods. Cell supernatants from the last day of the experiment were analyzed for IGFBP-1 protein, by ELISA. The values are mean ± SEM of three separate experiments done in triplicate. Significant differences, at P < 0.05, between groups, are designated by different lowercase letters. Note the decrease in IGFBP-1 secretion after the addition of IL-1ß to the dbcAMP and hormone-treated cells.

	Discussion

Top Abstract Introduction Materials and Methods Results Discussion References

 
Our previous studies in the baboon have clearly demonstrated that IGFBP-1 gene expression in the endometrium is a conceptus-mediated response (7). Subsequent studies in vitro established that IGFBP-1 gene expression in decidualizing stromal fibroblasts requires the presence of both hormones and dbcAMP (8). This induction is associated with a concomitant decrease of -SMA expression in vivo (20) and in vitro (8). Furthermore, our subsequent in vivo studies suggested that COX-2 expression in stromal fibroblasts, underlying the implantation site, precedes IGFBP-1 induction in the decidualized stromal cells (12).

Because IL-1ß is expressed both in the progestational endometrium and in trophoblast cells (23, 24), we evaluated IL-1ß as one possible factor that could influence differentiation of stromal cells into decidual cells. IL-1ß has been reported to be actively involved in fetal-maternal interactions (9, 10), but its role in decidualization has not been clarified. In addition, IL-1 can modulate changes in the cytoskeleton (25, 26) and induce COX-2 gene expression (13, 27). Our present data suggest that IL-1ß activates a signaling pathway that induces COX-2 gene expression followed by an increase in IGFBP-1 expression. These data are in agreement with our previous in vivo studies (7, 12). However, there are other reports suggesting that IL-1ß is inhibitory to decidualization (21, 22). In both these studies, the inhibitory effects of IL-1ß on IGFBP-1 and/or PRL synthesis were determined in the presence of dbcAMP or PGE₂. Our data are also in agreement with these previously published studies and confirm that IL-1ß in the presence of hormones inhibits dbcAMP-induced IGFBP-1 when all agents are added together.

The up-regulation of IGFBP-1, in response to IL-1ß, is in agreement with previous in vivo studies in rats and mice (27, 28). Intraperitoneal injections of IL-1 acutely increased serum levels of IGFBP-1 protein and hepatic IGFBP-1 mRNA synthesis within 2 h of stimulation (29). The direct effects of cytokines on IGFBP-1 induction were confirmed in vitro using HepG2 cells (29). IL-1 is a cytokine with many functions and is capable of inducing multiple immediate signaling pathways in the cells (reviewed in 30). We detected phosphorylated IB- protein after 10 min of IL-1ß treatment, suggesting possible activation of the NFB transcription factor in stromal fibroblasts. The other cascades activated by IL-1 are those activating the three best-known types of the MAPK subfamilies, namely p42/p44 extracellular signal-regulated protein kinase, c-Jun N-terminal kinase, p38 MAPK (31). In our studies we also detected activation of p38 MAPK kinase after IL-1ß treatment of baboon and human stromal cells.

IL-1 acts by inducing many genes (e.g. IL-2 and IL-6), chemokines (e.g. IL-8 and monocyte chemotactic protein 1), proteases (e.g. collagenase and stromelysin), adhesion molecules (e.g. intracellular adhesion molecule 1 and E-selectin), and cyclooxygenase (reviewed in 32). In our previous studies, COX-2 expression was detected specifically at the implantation site in baboons (13). In this report, we demonstrate that, in both baboon and human stromal cells cultured in vitro, treatment with IL-1ß induced COX-2 gene expression and PGE₂ synthesis. The IL-1ß-induced COX-2 activation contributes to the expression of IGFBP-1, because NS-398, a specific COX-2 inhibitor, partially inhibits the measurable IGFBP-1 protein levels in stromal cells. The lack of complete inhibition indicates that IL-1ß activates at least two immediate signaling pathways leading to IGFBP-1 synthesis in uterine stromal fibroblasts. The alternate pathway may be activated by IL-6. IL-1 has been shown to induce IL-6 production in cultured human decidual cells, and this induction can be prevented by actinomycin treatment (33). IL-6 elevates a 30- to 32-kDa IGF-binding protein in the plasma of mice and increases IGFBP-1 production in HepG2 cells (34). Moreover, specific regions on the IGFBP-1 promoter are regulated by IL-6 (35, 36). Thus, it is conceivable that activation of IL-6 in addition to COX-2 by IL-1ß could account for the lack of complete inhibition of IGFBP-1 expression in the presence of the COX-2 inhibitor. In fact, stimulation with IL-1ß induces IL-6 gene expression in HuF (Strakova, Hales and Fazleabas, unpublished results).

Our current studies have established the signaling pathways activated by IL-1ß in stromal fibroblasts and their role in inducing IGFBP-1 gene expression. The response to IL-1ß has a hormone-independent (COX-2 induction) and hormone-dependent (IGFBP-1 induction) pathway. In the presence of hormones, IL-1ß induces a similar pattern of IGFBP-1 expression and synthesis as that observed with hormones and dbcAMP (8). However, if dbcAMP is present together with IL-1ß, the differentiation process and IGFBP-1 induction are inhibited, suggesting the negative cross-talk between the two pathways. We can only hypothesize how the inhibition is accomplished. The response to IL-1ß or dbcAMP in the presence of hormones, in terms of IGFBP-1 expression, takes 2–3 days. This implies that, after initial stimulus, the decidualization process requires several intermediate steps. IL-1ß-induced PGE₂ causes additional cAMP release, because cAMP is a second messenger of PGE₂ stimulation (37). The cAMP is known to transmit its signals primarily through PKA, but expression of mRNA of PKA subunits did not differ in nondecidualized and decidualized cells (8). During the course of decidualization, expression of the inducible cAMP early repressor (ICER) is enhanced (38). The ICER is an isoform generated within the cAMP response element (CRE)-binding modulator (CREM) gene, which encodes both activators and repressors of cAMP-induced transcription. Thus, it is conceivable that, in the presence of both dbcAMP and IL-1ß, there is a premature increase in ICER expression that can subsequently inhibit the ability of PGE₂-induced cAMP to act positively and enhance IGFBP-1 transcription. Studies to test this hypothesis are currently underway in our laboratory.

Thus, it is evident that cytokines (IL-1) and mediators of cAMP activation (i.e. PGE₂) have independent stimulating effects, but synergistic inhibitory effects, in differentiating stromal fibroblasts. The complex signaling pathways activated during implantation may play a critical role in maintaining the appropriate homeostasis required for decidualization and trophoblast invasion.

	Acknowledgments

 
We thank Dr. Douglas Kniss (Ohio State University, Columbus, OH) for his kind gift of HES cell line, and Dr. David Sherwood (University of Illinois, Urbana-Champaign, IL) for purified porcine relaxin.

	Footnotes

 
¹ This work was supported by NIH Grant HD-36759.

Received March 13, 2000.

	References

Top Abstract Introduction Materials and Methods Results Discussion References

 

1. Fazleabas AT, Donnelly KM, Mavrogianis PA, Verhage HG 1993 Secretory and morphological changes in the baboon (Papio anubis) uterus and placenta during early pregnancy. Biol Reprod 49:695–704[Abstract]
2. Fazleabas AT, Donnelly KM, Sudha S, Fortman JD, Miller JB 1999 Modulation of the baboon (Papio anubis) uterine endometrium by chorionic gonadotropin during the period of uterine receptivity. Proc Natl Acad Sci USA 96:2543–2548[Abstract/Free Full Text]
3. Rutanen EM, Pekonen F, Makinen T 1988 Soluble 34K binding protein inhibits the binding of insulin-like growth factor I to its cell receptors in human secretory phase endometrium: evidence for autocrine/paracrine regulation of growth factor action. J Clin Endocrinol Metab 66:173–180[Abstract]
4. Fazleabas AT, Verhage HG, Waites G, Bell SC 1989 Characterization of an insulin-like growth factor binding protein (IGFBP), analogous to human pregnancy-associated secreted 1-globulin (1-PEG), in decidua of the baboon (Papio anubis) placenta. Biol Reprod 40:873–885[Abstract]
5. Fazleabas AT, Verhage HG, Bell SC 1989 Steroid-induced proteins of the primate oviduct and uterus: potential regulators of reproductive function. In: Krey LC, Guylas BJ, McCraken JA (eds) Autocrine and Paracrine Mechanisms in Reproductive Endocrinology. Plenum Publishing Corp., New York, pp 115–136
6. Hild-Petito S, Donnelly KM, Miller JB, Verhage HG, Fazleabas AT 1995 A baboon (Papio anubis) simulated-pregnant model: cell specific expression of insulin-like growth factor binding protein-1 (IGFBP-1), type I IGF receptor (IGF-1R) and retinol binding protein (RBP) in the uterus. Endocrine 3:639–651
7. Tarantino S, Verhage HG, Fazleabas AT 1992 Regulation of insulin-like growth factor binding proteins in the baboon (Papio anubis) uterus during early pregnancy. Endocrinology 130:2354–2362[Abstract]
8. Kim JJ, Jaffe RC, Fazleabas AT 1998 Comparative studies on the in vitro decidualization process in the baboon (Papio anubis) and human. Biol Reprod 59:160–168[Abstract/Free Full Text]
9. Simon C, Frances A, Piquette GN, Hendrickson M, Milki A, Polan ML 1994 Interleukin-1 system in the materno-trophoblast unit in human implantation: immunohistochemical evidence for autocrine/paracrine function. J Clin Endocrinol Metab 78:847–854[Abstract]
10. Simon C, Mercader A, Gimeno MJ, Pellicer A 1997 The interleukin-1 system and human implantation. Am J Reprod Immunol 37:64–72
11. Lim H, Paria B, Das SK, Dinchuk JE, Langenbach R, Trzaskos JM, Dey SK 1997 Multiple female reproductive failures in cyclooxygenase-2 deficient mice. Cell 91:197–208[CrossRef][Medline]
12. Kim JJ, Das SK, Wang J, Bambra C, Dey SK, Fazleabas AT 1999 Expression of cyclooxygenase-1 and -2 in the baboon endometrium during the menstrual cycle and pregnancy. Endocrinology 140:2672–2678[Abstract/Free Full Text]
13. Kniss DA, Zimmerman PD, Garver CL, Fertel RH 1997 Interleukin-1 receptor antagonist blocks interleukin-1-induced expression of cyclooxygenase-2 in endometrium. Am J Obstet Gynecol 177:559–567[CrossRef][Medline]
14. Huang JC, Liu DY, Yadollahi S, Wu KK, Dawood MY 1998 Interleukin-1ß induces cyclooxygenase-2 gene expression in cultured endometrial stromal cells. J Clin Endocrinol Metab 83:538–541[Abstract/Free Full Text]
15. Markoff E, Zeitler P, Peleg S, Handwerger S 1983 Characterization of the synthesis and release of prolactin by an enriched fraction of human decidual cells. J Clin Endocrinol Metab 56:962–968[Abstract]
16. Richards RA, Brar AK, Frank GR, Hartman SM, Jikihara H 1995 Fibroblast cells from term human decidua closely resemble endometrial stromal cells: induction of prolactin and insulin-like growth factor binding protein-1 expression. Biol Reprod 52:609–615[Abstract]
17. Strakova Z, Copland JA, Lolait SG, Soloff MS 1998 ERK-2 mediates oxytocin-stimulated PGE₂ synthesis. Am J Physiol 274:E634–E641
18. Baldwin Jr AS 1996 The NF-B and IB proteins: new discoveries and insights. Annu Rev Immunol 14:649–683[CrossRef][Medline]
19. Chen Z, Hagler J, Palombella VJ, Melandri F, Scherer D, Ballard D, Maniatis T 1995 Signal-induced site-specific phosphorylation targets IB to the ubiquitin-proteasome pathway. Genes Dev 9:1586–1597[Abstract/Free Full Text]
20. Christensen S, Verhage HG, Nowak G, de Lanerolle P, Fleming S, Bell SC, Fazleabas AT, Hild-Petito S 1995 Smooth muscle myosin II and smooth muscle actin expression in the baboon (Papio anubis) uterus is associated with glandular secretory activity and stromal cell transformation. Biol Reprod 53:596–606
21. Frank GR, Brar AK, Jikihara H, Cedars MI, Handwerger S 1995 Interleukin-1ß and the endometrium: an inhibitor of stromal cell differentiation and possible autoregulator of decidualization in humans. Biol Reprod 52:184–191[Abstract]
22. Mizuno M, Tanaka T, Umesaki N, Ogita S 1999 Inhibition of cAMP-mediated decidualization in human endometrial cells by IL-1ß and laminin. Horm Metab Res 31:307–310[Medline]
23. Kauma S, Matt D, Strom S, Eierman D, Tuner T 1990 Interleukin 1-ß, human leukocyte antigen HLA-DR , and transforming growth factor-ß expression in endometrium, placenta and placental membranes. Am J Obstet Gynecol 163:1430–1437[Medline]
24. De los Santos MJ, Mercader A, Frances A, Portoles E, Remohi J, Pellicer A, Simon C 1996 Role of endometrial factors in regulating secretion of components of the immunoreactive human embryonic interleukin-1 system during embryonic development. Biol Reprod 54:563–574[Abstract]
25. Singh R, Wang B, Shirvaikar A, Khan S, Kamat S, Schelling JR, Konieczkowski M, Sedor JR 1999 The IL-1 receptor and Rho directly associate to drive cell activation in inflammation. J Clin Invest 103:1561–1570[Medline]
26. Zhu P, Xiong W, Rodgers G, Qwarnstrom EE 1998 Regulation of interleukin 1 signalling through integrin binding and actin reorganization: disparate effects on NF-B and stress kinase pathways. Biochem J 330:975–981
27. Guan Z, Buckman SY, Miller BW, Springer LD, Morrison AR 1998 Interleukin-1ß-induced cyclooxygenase-2 expression requires activation of both c-Jun NH₂ terminal kinase and p38 MAPK signal pathways in rat mesengial cells. J Biol Chem 273:28670–28676[Abstract/Free Full Text]
28. Fan J, Wojnar MM, Theodorakis M, Lang CH 1996 Regulation of insulin-like growth factor (IGF)-I mRNA and peptide and IGF-binding proteins by interleukin-1 Am J Physiol 270:R621–R629
29. Benbassat CA, Lazarus DD, Cichy SB, Evans TM, Moldawer LL, Lowry SF, Unterman TG 1999 Interleukin-1 (IL-1) and tumor necrosis factor (TNF) regulate insulin-like growth factor binding protein-1 (IGFBP-1) levels and mRNA abundance in vivo and in vitro. Horm Metab Res 31:209–215[Medline]
30. O’Neill LA, Greene C 1998 Signal transduction pathways activated by the IL-1 receptor family: ancient signaling machinery in mammals, insects, and plants. J Leukoc Biol 63:650–657[Abstract]
31. Cano E, Mahadevan LC 1995 Parallel signal processing among mammalian MAPKs. Trends Biochem Sci 20:117–122[CrossRef][Medline]
32. Dinarello CA 1996 Biologic basis for interleukin-1 in disease. Blood 87:2095–2147[Abstract/Free Full Text]
33. Dudley DJ, Trautman MS, Araneo BA, Edwin SS, Mitchell MD 1992 Decidual cell biosynthesis of interleukin-6:regulation by inflammatory cytokines. J Clin Endocrinol Metab 74:884–889[Abstract]
34. Samstein B, Hoimes ML, Fan J, Frost RA, Gelato MC, Lang CH 1996 IL-6 stimulation of insulin-like growth factor binding protein (IGFBP)-1 production. Biochem Biophys Res Commun 228:611–615[CrossRef][Medline]
35. Cressman DE, Greenbaum LE, De Angelis RA, Cilberto G, Furth EE, Poli V, Taub R 1996 Liver failure and defective hepatocyte regeneration in interleukin-6-deficient mice. Science 22:1379–1383
36. Crissey MA, Leu JI, De Angelis RA, Greenbaum LE, Scearce M, Kovalovich K, Taub R 1999 Liver-specific and proliferation-induced deoxyribonuclease I hypersensitive sites in the mouse insulin-like growth factor binding protein-1 gene. Hepatology 30:1187–1197[CrossRef][Medline]
37. Yee GM, Kennedy TG 1991 Role of cyclic adenosine 3', 5'-monophosphate in mediating the effect of prostaglandin E2 on decidualization in vitro. Biol Reprod 45:163–171[Abstract]
38. Gellersen B, Kempf R, Telgmann R 1997 Human endometrial stromal cells express novel isoforms of the transcriptional modulator CREM and up-regulate ICER in the course of decidualization. Mol Endocrinol 11:97–113[Abstract/Free Full Text]

This article has been cited by other articles:

				S. S D'Souza, A. T Fazleabas, P. Banerjee, J. R. A Sherwin, A. M Sharkey, M. C Farach-Carson, and D. D Carson Decidual Heparanase Activity Is Increased During Pregnancy in the Baboon (Papio anubis) and in In Vitro Decidualization of Human Stromal Cells Biol Reprod, February 1, 2008; 78(2): 316 - 323. [Abstract] [Full Text] [PDF]

				I. Ihnatovych, W. Hu, J. L. Martin, A. T. Fazleabas, P. de Lanerolle, and Z. Strakova Increased Phosphorylation of Myosin Light Chain Prevents in Vitro Decidualization Endocrinology, July 1, 2007; 148(7): 3176 - 3184. [Abstract] [Full Text] [PDF]

				J. R. A. Sherwin, A. M. Sharkey, P. Cameo, P. M. Mavrogianis, R. D. Catalano, S. Edassery, and A. T. Fazleabas Identification of Novel Genes Regulated by Chorionic Gonadotropin in Baboon Endometrium during the Window of Implantation Endocrinology, February 1, 2007; 148(2): 618 - 626. [Abstract] [Full Text] [PDF]

				B. Gellersen, J. Briese, M. Oberndorfer, K. Redlin, A. Samalecos, D.-U. Richter, T. Loning, H.-M. Schulte, and A.-M. Bamberger Expression of the Metastasis Suppressor KAI1 in Decidual Cells at the Human Maternal-Fetal Interface: Regulation and Functional Implications Am. J. Pathol., January 1, 2007; 170(1): 126 - 139. [Abstract] [Full Text] [PDF]

				P. Cameo, M. Szmidt, Z. Strakova, P. Mavrogianis, K. L. Sharpe-Timms, and A. T. Fazleabas Decidualization Regulates the Expression of the Endometrial Chorionic Gonadotropin Receptor in the Primate Biol Reprod, November 1, 2006; 75(5): 681 - 689. [Abstract] [Full Text] [PDF]

				A. Jasinska, Z. Strakova, M. Szmidt, and A. T. Fazleabas Human Chorionic Gonadotropin and Decidualization in Vitro Inhibits Cytochalasin-D-Induced Apoptosis in Cultured Endometrial Stromal Fibroblasts Endocrinology, September 1, 2006; 147(9): 4112 - 4121. [Abstract] [Full Text] [PDF]

				A. G. Braundmeier, A. T. Fazleabas, B. A. Lessey, H. Guo, B. P. Toole, and R. A. Nowak Extracellular Matrix Metalloproteinase Inducer Regulates Metalloproteinases in Human Uterine Endometrium J. Clin. Endocrinol. Metab., June 1, 2006; 91(6): 2358 - 2365. [Abstract] [Full Text] [PDF]

				M. Tang, A. Mikhailik, I. Pauli, L. C. Giudice, A. T. Fazelabas, S. Tulac, D. D. Carson, D. G. Kaufman, C. Barbier, J. W. M. Creemers, et al. Decidual Differentiation of Stromal Cells Promotes Proprotein Convertase 5/6 Expression and Lefty Processing Endocrinology, December 1, 2005; 146(12): 5313 - 5320. [Abstract] [Full Text] [PDF]

				J.J. Kim, O.L. Buzzio, S. Li, and Z. Lu Role of FOXO1A in the Regulation of Insulin-Like Growth Factor-Binding Protein-1 in Human Endometrial Cells: Interaction with Progesterone Receptor Biol Reprod, October 1, 2005; 73(4): 833 - 839. [Abstract] [Full Text] [PDF]

				Z. Strakova, P. Mavrogianis, X. Meng, J. M. Hastings, K. S. Jackson, P. Cameo, A. Brudney, O. Knight, and A. T. Fazleabas In Vivo Infusion of Interleukin-1{beta} and Chorionic Gonadotropin Induces Endometrial Changes that Mimic Early Pregnancy Events in the Baboon Endocrinology, September 1, 2005; 146(9): 4097 - 4104. [Abstract] [Full Text] [PDF]

				C.A. White, E. Dimitriadis, A.M. Sharkey, and L.A. Salamonsen Interleukin-11 inhibits expression of insulin-like growth factor binding protein-5 mRNA in decidualizing human endometrial stromal cells Mol. Hum. Reprod., September 1, 2005; 11(9): 649 - 658. [Abstract] [Full Text] [PDF]

				G. Nie, Y. Li, M. Wang, Y. X. Liu, J. K. Findlay, and L. A. Salamonsen Inhibiting Uterine PC6 Blocks Embryo Implantation: An Obligatory Role for a Proprotein Convertase in Fertility Biol Reprod, April 1, 2005; 72(4): 1029 - 1036. [Abstract] [Full Text] [PDF]

				A. Jasinska, V. Han, A. T. Fazleabas, and J. J. Kim Induction of Insulin-Like Growth Factor Binding Protein-1 Expression in Baboon Endometrial Stromal Cells by Cells of Trophoblast Origin Reproductive Sciences, September 1, 2004; 11(6): 399 - 405. [Abstract] [PDF]

				Z. Strakova, M. Szmidt, S. Srisuparp, and A. T. Fazleabas Inhibition of Matrix Metalloproteinases Prevents the Synthesis of Insulin-Like Growth Factor Binding Protein-1 during Decidualization in the Baboon Endocrinology, December 1, 2003; 144(12): 5339 - 5346. [Abstract] [Full Text] [PDF]

				H.-Y. Li, S.-P. Chang, C.-C. Yuan, H.-T. Chao, H.-T. Ng, and Y.-J. Sung Induction of p38 Mitogen-Activated Protein Kinase-Mediated Apoptosis Is Involved in Outgrowth of Trophoblast Cells on Endometrial Epithelial Cells in a Model of Human Trophoblast-Endometrial Interactions Biol Reprod, November 1, 2003; 69(5): 1515 - 1524. [Abstract] [Full Text] [PDF]

				K. T. Coschigano, A. N. Holland, M. E. Riders, E. O. List, A. Flyvbjerg, and J. J. Kopchick Deletion, But Not Antagonism, of the Mouse Growth Hormone Receptor Results in Severely Decreased Body Weights, Insulin, and Insulin-Like Growth Factor I Levels and Increased Life Span Endocrinology, September 1, 2003; 144(9): 3799 - 3810. [Abstract] [Full Text] [PDF]

				J. J. Kim, H. S. Taylor, G. E. Akbas, I. Foucher, A. Trembleau, R. C. Jaffe, A. T. Fazleabas, and T. G. Unterman Regulation of Insulin-Like Growth Factor Binding Protein-1 Promoter Activity by FKHR and HOXA10 in Primate Endometrial Cells Biol Reprod, January 1, 2003; 68(1): 24 - 30. [Abstract] [Full Text] [PDF]

				I. Foucher, M. Volovitch, M. Frain, J. J. Kim, J.-C. Souberbielle, L. Gan, T. G. Unterman, A. Prochiantz, and A. Trembleau Hoxa5 overexpression correlates with IGFBP1 upregulation and postnatal dwarfism: evidence for an interaction between Hoxa5 and Forkhead box transcription factors Development, September 1, 2002; 129(17): 4065 - 4074. [Abstract] [Full Text] [PDF]

				L. C. Kao, S. Tulac, S. Lobo, B. Imani, J. P. Yang, A. Germeyer, K. Osteen, R. N. Taylor, B. A. Lessey, and L. C. Giudice Global Gene Profiling in Human Endometrium during the Window of Implantation Endocrinology, June 1, 2002; 143(6): 2119 - 2138. [Abstract] [Full Text] [PDF]

This Article Abstract Full Text (PDF)