Nowak Karolina, Jabłońska Ewa, Garley Marzena, Radziwon Piotr, Ratajczak-Wrona Wioletta
Department of Immunology, Medical University of Bialystok, Poland.
Department of Immunology, Medical University of Bialystok, Poland.
Mol Cell Endocrinol. 2021 Dec 1;538:111470. doi: 10.1016/j.mce.2021.111470. Epub 2021 Oct 1.
Parabens, including the most common methylparaben (MeP), are popular preservatives, which possess estrogenic activity. The aims of this study were to assess the impact of MeP on estrogen receptors (ERs) and/or NF-κB-dependent generation of IL-8 and production of nitric oxide (NO), and also to verify the hypothesis about the crosstalk of ERs with NF-κB in xenoestrogen-exposed neutrophils. Human neutrophils were incubated for 20-h with MeP (0.06 μM) and/or ER antagonist (1 μM) and/or NF-κB inhibitor (100 μM). After the isolation of cell lysates and cytoplasmic and nuclear fraction, the expression of ERα, ERβ, p-IKKα/β, p65 NF-κB, and inducible nitric oxide synthase (iNOS) was measured by Western blot analysis, The concentration of NO was evaluated by Griess reaction, and that of IL-8 was measured by ELISA. The results showed that MeP modulated the expression of ERα, but not ERβ. Exposure to paraben activated iKKα/β-dependent NF-κB pathway, but translocation of p65 NF-κB into the cell nucleus was inhibited by ERs. MeP also decreased the iNOS-dependent production of NO, but did not influence the secretion of IL-8 by neutrophils. The study indicates that MeP may affect the functioning of human neutrophils by modulating intracellular signal transduction pathways, including ERs and NF-κB pathway.
对羟基苯甲酸酯，包括最常见的对羟基苯甲酸甲酯（MeP），是常用的防腐剂，具有雌激素活性。本研究的目的是评估MeP对雌激素受体（ERs）和/或NF-κB依赖的白细胞介素-8生成及一氧化氮（NO）产生的影响，并验证在暴露于外源性雌激素的中性粒细胞中ERs与NF-κB相互作用的假设。将人类中性粒细胞与MeP（0.06 μM）和/或ER拮抗剂（1 μM）和/或NF-κB抑制剂（100 μM）孵育20小时。分离细胞裂解物、细胞质和细胞核组分后，通过蛋白质印迹分析测定ERα、ERβ、磷酸化IKKα/β、p65 NF-κB和诱导型一氧化氮合酶（iNOS）的表达，通过格里斯反应评估NO的浓度，通过酶联免疫吸附测定法测量IL-8的浓度。结果表明，MeP调节ERα的表达，但不调节ERβ的表达。暴露于对羟基苯甲酸酯激活了iKKα/β依赖的NF-κB途径，但ERs抑制了p65 NF-κB向细胞核的转位。MeP还降低了iNOS依赖的NO产生，但不影响中性粒细胞分泌IL-8。该研究表明，MeP可能通过调节包括ERs和NF-κB途径在内的细胞内信号转导途径来影响人类中性粒细胞的功能。
