Alvarado-Díaz Carlos Patricio, Núñez Marco Tulio, Devoto Luigi, González-Ramos Reinaldo
Maternal and Child Research Institute, Department of Obstetrics and Gynecology, Faculty of Medicine, San Borja Arriarán Hospital, University of Chile, Santiago, Chile.
Cellular Dynamics and Biotechnology Institute, Faculty of Sciences, University of Chile, Santiago, Chile.
Fertil Steril. 2015 Feb;103(2):439-47. doi: 10.1016/j.fertnstert.2014.10.046. Epub 2014 Dec 12.
To evaluate the effect of iron overload on nuclear factor kappa-B (NF-κB) activation in human endometrial stromal cells (ESCs).
Experimental study.
University 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.
评估铁过载对人子宫内膜基质细胞（ESCs）中核因子κB（NF-κB）激活的影响。
实验研究。
大学医院研究实验室。
10名健康女性。
从子宫内膜活检中分离出的ESCs与50μM硫酸亚铁（FeSO₄）或赋形剂一起孵育。使用抑制IKKβ（IκBα（NF-κB的抑制蛋白）的激酶）的NF-κB抑制剂[5-（对氟苯基）-2-脲基]噻吩-3-甲酰胺（TPCA-1）来预防铁过载刺激的ESCs中NF-κB的变化。
通过p65：DNA结合活性免疫检测法评估NF-κB激活。通过蛋白质免疫印迹法评估IκBα、p65和细胞间黏附分子（ICAM）-1蛋白的表达。使用条件培养基通过酶联免疫吸附测定法（ELISA）测量ESC可溶性ICAM（sICAM）-1的分泌。
与基础状态相比，孵育30分钟后，铁过载增加了ESCs中p65：DNA结合活性，并降低了IκBα和p65的细胞质表达。铁过载处理24小时后，ESC ICAM-1的表达和sICAM-1的分泌高于未处理时。TPCA-1可预防铁过载诱导的p65：DNA结合增加和IκBα降解。
铁过载激活ESCs中的IKKβ，刺激NF-κB途径并增加ICAM-1表达和sICAM-1分泌。这些结果表明，铁过载在健康的ESCs上诱导了一种前子宫内膜异位症表型，这可能参与子宫内膜异位症的发病机制和发展。
