Department of Immunology, Medical University of Bialystok, Waszyngtona 15A, 15-269, Bialystok, Poland.
Regional Centre for Transfusion Medicine, Bialystok, Poland.
Environ Sci Pollut Res Int. 2020 Feb;27(6):6540-6548. doi: 10.1007/s11356-019-07388-w. Epub 2019 Dec 23.
This study was conducted to verify a hypothesis that immune cells are a target for the action of endocrine disrupting chemicals (EDCs) by investigating whether methylparaben (MeP) modulates human neutrophil functions. Neutrophils isolated from 15 donor samples were studied. Cells were incubated in the presence of increasing MeP concentrations (0.06, 0.8, 10, and 20 μM). The cytotoxic effect of MeP on neutrophils was evaluated by the MTT test. The ability of the neutrophils for chemotaxis, phagocytosis, NADPH oxidase activity, and superoxide anion generation was assessed in Boyden's chamber, Park's method with latex, the NBT test, and the cytochrome C reduction test, respectively. The total nitric oxide (NO) concentration was measured by the Griess reaction. There was no observable cytotoxic effect of MeP on human neutrophils. MeP (10 and 20 μM) exposure decreased neutrophilic ability for the tested functions, except for NO production. In neutrophils incubated with MeP (0.8 μM as well as 0.06 and 0.8 μM, respectively), we observed a decreased activity of NADPH oxidase as well as decreased generation of superoxide anion. Our results suggest a suppressive effect of MeP on the tested functions of human neutrophils, which confirms that immune cells are vulnerable to EDC action. Therefore, the disturbance of neutrophils' oxygen-dependent phagocytic function as a result of exposure to environmental doses of MeP action could lead to impairment of innate immune responses in humans exposed to xenoestrogens.
本研究旨在验证一种假设,即免疫细胞是内分泌干扰化学物质(EDCs)作用的靶标,通过研究是否对羟基苯甲酸甲酯(MeP)调节人中性粒细胞功能。从 15 个供体样本中分离出中性粒细胞进行研究。将细胞在不同浓度的 MeP(0.06、0.8、10 和 20 μM)存在下孵育。通过 MTT 试验评估 MeP 对中性粒细胞的细胞毒性作用。通过 Boyden 室、Park 法与乳胶、NBT 试验和细胞色素 C 还原试验分别评估中性粒细胞的趋化性、吞噬作用、NADPH 氧化酶活性和超氧阴离子生成能力。通过 Griess 反应测定总一氧化氮(NO)浓度。MeP 对人中性粒细胞无明显的细胞毒性作用。MeP(10 和 20 μM)暴露降低了中性粒细胞的测试功能,除了 NO 产生。在与 MeP(0.8 μM 以及 0.06 和 0.8 μM 分别孵育的中性粒细胞中),我们观察到 NADPH 氧化酶活性降低,超氧阴离子生成减少。我们的结果表明 MeP 对人中性粒细胞的测试功能具有抑制作用,这证实了免疫细胞易受 EDC 作用的影响。因此,暴露于环境剂量的 MeP 作用会干扰中性粒细胞的氧依赖性吞噬功能,从而导致接触外源性雌激素的人类先天免疫反应受损。