Iron overload–modulated nuclear factor kappa-B activation in human endometrial stromal cells as a mechanism postulated in endometriosis pathogenesis

Carlos Patricio Alvarado-Díaz, Alvarado-Díaz CP, Marco T. Núñez, Marco Tulio Núñez, Núñez MT, Luigi Devoto, Devoto L, González-Ramos R, Reinaldo González-Ramos
Fertility and sterility · 2014 · vol. 103(2) , pp. 439–447 · doi:10.1016/j.fertnstert.2014.10.046 · PMID:25500022 · W2072916560
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Iron overload activates the NF-kappa B pathway in human endometrial stromal cells, increasing ICAM-1 expression and secretion, suggesting a role in endometriosis pathogenesis.

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This experimental study evaluated how iron overload affects nuclear factor kappa-B (NF-κB) activation in human endometrial stromal cells (ESCs) isolated from endometrial biopsies of ten healthy women. ESCs were incubated with 50 μM FeSO4 (or vehicle) and NF-κB pathway activation was assessed by p65 DNA-binding assays, Western blot measurements of IκBα, p65, and intercellular adhesion molecule-1 (ICAM-1), and ELISA measurement of soluble ICAM-1 secretion after conditioned-medium collection. Iron overload increased p65 DNA-binding activity and decreased cytoplasmic IκBα and p65 at 30 minutes, and increased ICAM-1 expression and sICAM-1 secretion at 24 hours, while the IKKβ inhibitor TPCA-1 prevented these iron-overload–induced NF-κB changes. The study is limited by its in vitro design using cells from healthy women, and it does not directly test endometriosis in patients. This paper is centrally about endometriosis—proposes that iron overload activates NF-κB signaling in endometrial stromal cells, promoting a proendometriotic phenotype relevant to endometriosis pathogenesis.

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Abstract

ObjectiveTo evaluate the effect of iron overload on nuclear factor kappa-B (NF-κB) activation in human endometrial stromal cells (ESCs).DesignExperimental study.SettingUniversity hospital research laboratory.Patient(s)Ten healthy women.Intervention(s)Isolated ESCs from endometrial biopsies were incubated with 50 μM FeSO(4) or vehicle. The NF-κB inhibitor [5-(p-fluorophenyl)-2-ureido] thiophene-3-carboxamide (TPCA-1), which inhibits IKKβ, the kinase of IκBα (inhibitory protein of NF-κB), was used to prevent iron overload-stimulated NF-κB changes in ESCs.Main outcome measure(s)NF-κB activation was assessed by p65:DNA-binding activity immunodetection assay. IκBα, p65, and intercellular adhesion molecule (ICAM)-1 proteins expression was evaluated by Western blots. ESC soluble ICAM (sICAM)-1 secretion was measured by ELISA using conditioned medium.Result(s)Iron overload increased p65:DNA-binding activity and decreased IκBα and p65 cytoplasmic expression in ESCs after 30 minutes of incubation as compared with the basal condition. ESC ICAM-1 expression and sICAM-1 secretion were higher after 24 hours of iron overload treatment than in the absence of treatment. TPCA-1 prevented the iron overload-induced increase of p65:DNA binding and IκBα degradation.Conclusion(s)Iron overload activates IKKβ in ESCs, stimulating the NF-κB pathway and increasing ICAM-1 expression and sICAM-1 secretion. These results suggest that iron overload induces a proendometriotic phenotype on healthy ESCs, which could participate in endometriosis pathogenesis and development.
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Iron overload-modulated nuclear factor kappa-B activation in human endometrial stromal cells as a mechanism postulated in endometriosis pathogenesis Artículo Open/ Download Access note Acceso abierto Publication date 2015Metadata Show full item record Cómo citar Alvarado Díaz, Carlos Patricio Cómo citar Iron overload-modulated nuclear factor kappa-B activation in human endometrial stromal cells as a mechanism postulated in endometriosis pathogenesis Author Abstract Objective: To evaluate the effect of iron overload on nuclear factor kappa-B (NF-kappa B) activation in human endometrial stromal cells (ESCs). Design: Experimental study. Setting: University hospital research laboratory. Patient(s): Ten healthy women. Intervention(s): Isolated ESCs from endometrial biopsies were incubated with 50 mu M FeSO4 or vehicle. The NF-kappa B inhibitor [5-(p-fluorophenyl)-2-ureido] thiophene-3-carboxamide (TPCA-1), which inhibits IKK beta, the kinase of I kappa B alpha (inhibitory protein of NF-kappa B), was used to prevent iron overload-stimulated NP-kappa B changes in ESCs. Main Outcome Measure(s): NF-kappa B activation was assessed by p65:DNA-binding activity immunodetection assay. I kappa B alpha, p65, and intercellular adhesion molecule (ICAM)-1 proteins expression was evaluated by Western blots. ESC soluble ICAM (sICAM)-1 secretion was measured by ELISA using conditioned medium. Result(s): Iron overload increased p65:DNA-binding activity and decreased I kappa B alpha and p65 cytoplasmic expression in ESCs after 30 minutes of incubation as compared with the basal condition. ESC ICAM-1 expression and sICAM-1 secretion were higher after 24 hours of iron overload treatment than in the absence of treatment. TPCA-1 prevented the iron overload-induced increase of p65:DNA binding and I kappa B alpha degradation. Conclusion(s): Iron overload activates IKK beta in ESCs, stimulating the NF-kappa B pathway and increasing ICAM-1 expression and sICAM-1 secretion. These results suggest that iron overload induces a proendometriotic phenotype on healthy ESCs, which could participate in endometriosis pathogenesis and development. Indexation Artículo de publicación SCOPUS Artículo de publicación WoS Identifier URI: https://repositorio.uchile.cl/handle/2250/166370 DOI: 10.1016/j.fertnstert.2014.10.046 ISSN: 15565653 00150282 Quote Item Fertility and Sterility Vol. 103, No. 2, February 2015 Collections

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endometriosis

MeSH descriptors

Endometriosis Endometrium Endometrium Iron Overload NF-kappa B Adult Amides Amides Cells, Cultured Endometriosis Endometriosis Endometrium Endometrium Female Ferric Compounds Ferric Compounds Humans Iron Overload Iron Overload NF-kappa B

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