Fractalkine Regulates HEC-1A/JEG-3 Interaction by Influencing the Expression of Implantation-Related Genes in an In Vitro Co-Culture Model

Abstract

Embryo implantation is a complex process regulated by a network of biological molecules. Recently, it has been described that fractalkine (CX3CL1, FKN) might have an important role in the feto–maternal interaction during gestation since the trophoblast cells express fractalkine receptor (CX3CR1) and the endometrium cells secrete fractalkine. CX3CR1 controls three major signalling pathways, PLC-PKC pathway, PI3K/AKT/NFκB pathway and Ras-mitogen-activated protein kinases (MAPK) pathways regulating proliferation, growth, migration and apoptosis. In this study, we focused on the molecular mechanisms of FKN treatment influencing the expression of implantation-related genes in trophoblast cells (JEG-3) both in mono-and in co-culture models. Our results reveal that FKN acted in a concentration and time dependent manner on JEG-3 cells. FKN seemed to operate as a positive regulator of implantation via changing the action of progesterone receptor (PR), activin receptor and bone morphogenetic protein receptor (BMPR). FKN modified also the expression of matrix metalloproteinase 2 and 9 controlling invasion. The presence of HEC-1A endometrial cells in the co-culture contributed to the effect of fractalkine on JEG-3 cells regulating implantation. The results suggest that FKN may contribute to the successful attachment and implantation of embryo.

Three chemotypes of thyme (Thymus vulgaris L.) essential oil and their main compounds affect differently the IL-6 and TNFα cytokine secretions of BV-2 microglia by modulating the NF-κB and C/EBPβ signalling pathways

“…After the electrophoresis the protein content of the gels were transferred by electro- blotting to nitrocellulose membranes (Pall AG, Basel, Switzerland). The membranes were blocked with 5% non-fat dry milk in TBST (Tris buffer saline, 0.1% Tween-20) for 1 h at room temperature [ 33 ]. After the blocking step, the membranes were probed with the following polyclonal rabbit antibodies for overnight at 4 °C according to the manufacturer’s protocols: anti-NF-κB/p50 IgG (1:1000, Sigma-Aldrich Kft., Budapest, Hungary), anti-NF-κB/p65 IgG (1:2000, Cell Signaling Technology Europe, Leiden, The Netherlands) and anti-phospho-C/EBPβ IgG (1:1000, Thermo Fisher Scientific Inc., Waltham, MA).…”

“…Statistical analysis was performed using SPSS software (IBM Corporation, Armonk, NY, USA). Statistical significance was determined by Kruskal-Wallis one-way ANOVA non-parametric test using pairwise comparisons [ 33 ]. Data are shown as mean ± standard deviation (SD).…”

Immunomodulatory and Antiprotozoal Potential of Fabricated Sesamum radiatum Oil/Polyvinylpyrrolidone/Au Polymeric Bionanocomposite Film

“…The protein content of the gels was moved to nitrocellulose membranes (Pall AG, Basel, Switzerland) by electro-blotting after the electrophoresis. Then, 5% non-fat dry milk in tris buffer saline (TBST, 0.1% Tween-20) was used to block the membranes at room temperature for 1 h [39]. Then, the membranes were explored with polyclonal rabbit antibodies at 4 • C overnight, according to manufacturer's protocols: anti-NF-κB/p65 IgG (1:2000, Cell Signaling Technology Europe, Leiden, The Netherlands), anti-NF-κB/p50 IgG (1:1000, Sigma-Aldrich Kft., Budapest, Hungary), and anti-phospho-C/EBPβ IgG (1:1000, Thermo Fisher Scientific Inc., Waltham, MA, USA).…”

Modulatory Effects of Fractalkine on Inflammatory Response and Iron Metabolism of Lipopolysaccharide and Lipoteichoic Acid-Activated THP-1 Macrophages

“…According to the literature, fractalkine (CX3CL1) acts as a chemokine as well as an inflammatory mediator and a regulator of the iron metabolism of various cells [ 8 , 9 , 10 ]. Fractalkine is expressed by many types of cells (e.g., neurons, astrocytes, endometrial cells and immune cells) and is also present on the surface of the endothelial cells of the blood vessels as a membrane-bound form, which provides an anchoring site for the macrophages and lymphocytes [ 11 , 12 , 13 , 14 , 15 , 16 ]. Fractalkine also can be secreted as a soluble form by the proteolytic cleavage of the membrane-bound fractalkine acting as an inflammatory mediator [ 17 , 18 ].…”