Effects of subchronic exposure of nonylphenol on the expression of immune-related factors and estrogen receptors in the spleen of rats

IF 5.9 3区 环境科学与生态学 Q1 Environmental Science
Xiangjun Fu, Jie Xu, Chengyu Ni, Degang Yu, Haibo Wang, Pan Wang, Man Luo, Jie Yu
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引用次数: 2

Abstract

Background

Previous studies have shown that EDCs may activate nuclear transcription factor, such as activator protein-1 (AP-1), nuclear factor of activated Tcells (NF-AT) and nuclear factor kappa B (NF-κB) in the process of immune damage. At the same time, some experts believed that estrogen may play an important role in this process. As a typical representative of EDCs, nonylphenol (NP) has not been reported. The aim of this work was to explore the relationship between the immune inflammatory damage and the changes in estrogen expression in male rats during the chronic exposure to NP at environmental concentrations. Sixty SPF Sprague–Dawley rats were divided into five groups (n = 12 per group): blank control group (corn oil), low-dose NP exposure group (0.4 mg/kg/d), medium-dose NP exposure group (4 mg/kg/d), high-dose NP exposure group (40 mg/kg/d), and estradiol control group (E2: 30 μg/kg/d).

Results

Compared with the control group, rat spleen organ coefficient, number of spleen nodules, relative area of lymph nodes and white pulp were relatively reduced in the L (NP, 0.4 mg/kg) and H (NP, 40 mg/kg) exposure dose groups (P < 0.001). Lymphocytes were rich in cytoplasm, mitochondria were swollen, part of the cristae was reduced, and rough endoplasmic reticulum was expanded. The serum levels of IgG (P < 0.001) and IgM (P = 0.002) showed a downward trend. The percentage of Th cells (CD3+CD4+) was significantly decreased (P < 0.001), and the percentage of B lymphocytes shows an opposite trend (P < 0.001). Giemsa staining showed that the number of neutrophils (P < 0.001) was increased. The expressions of estrogen receptor ER-α and ER-β protein in the spleen increased significantly (P < 0.001). The expressions of AP-1 protein and NF-AT protein in the spleen were increased, and the expression of NF-KB protein was decreased (P < 0.001). The expressions of IL-4, ER-α and ER-β (P < 0.001) levels in serum increased. The mRNA-seq bioinformatics detection showed the final differentially expressed immune-inflammatory-related genes between the control and H-NP groups as follow: down-regulated: TLR4, Gata3, IL12, up-regulated: TNF-a, IL10, INOS. The mRNA expressions of ER-α, ER-β, NF-KB, IL4, AP-1, TLR4, Gata3, and NF-AT were consistent with the results of mRNA-seq analysis. NP content was correlated with the expressions of ER-α, ER-β, IL4, AP-1, NF-AT, TLR4, NF-KB, as well as IL-12 proteins in the spleen tissue ([r] < 1, P < 0.05).

Conclusions

Chronic exposure to NP at environmental concentration could cause immune dysfunction, resulting in immunotoxicity and inflammatory effects, and lead to changes in the activity of transcription factors and differential immune inflammatory factors in rats.

Graphical Abstract

壬基酚亚慢性暴露对大鼠脾脏免疫相关因子及雌激素受体表达的影响
背景已有研究表明,EDCs在免疫损伤过程中可能激活核转录因子,如活化蛋白-1 (AP-1)、活化t细胞核因子(NF- at)和核因子κB (NF-κB)。与此同时,一些专家认为雌激素可能在这一过程中起着重要作用。壬基酚(NP)作为EDCs的典型代表尚未见报道。本研究旨在探讨环境浓度NP对雄性大鼠免疫炎症损伤与雌激素表达变化的关系。将60只SPF级Sprague-Dawley大鼠分为空白对照组(玉米油)、低剂量NP暴露组(0.4 mg/kg/d)、中剂量NP暴露组(4 mg/kg/d)、高剂量NP暴露组(40 mg/kg/d)和雌二醇对照组(E2: 30 μg/kg/d) 5组,每组12只。结果与对照组相比,L (NP, 0.4 mg/kg)和H (NP, 40 mg/kg)暴露剂量组大鼠脾脏脏器系数、脾脏结节数量、淋巴结相对面积和白髓相对减少(P < 0.001)。细胞质中淋巴细胞丰富,线粒体肿胀,部分嵴缩小,粗面内质网扩张。血清IgG (P < 0.001)和IgM (P = 0.002)水平呈下降趋势。Th细胞(CD3+CD4+)百分比显著降低(P < 0.001), B淋巴细胞百分比呈相反趋势(P < 0.001)。吉姆萨染色显示中性粒细胞数量增加(P < 0.001)。脾脏雌激素受体ER-α和ER-β蛋白表达显著升高(P < 0.001)。脾脏AP-1蛋白和NF-AT蛋白表达升高,NF-KB蛋白表达降低(P < 0.001)。血清IL-4、ER-α、ER-β表达升高(P < 0.001)。mRNA-seq生物信息学检测显示,对照组与H-NP组最终差异表达的免疫炎症相关基因如下:下调:TLR4、Gata3、IL12,上调:TNF-a、IL10、INOS。ER-α、ER-β、NF-KB、IL4、AP-1、TLR4、Gata3、NF-AT mRNA表达与mRNA-seq分析结果一致。NP含量与脾组织中ER-α、ER-β、il - 4、AP-1、NF-AT、TLR4、NF-KB、IL-12蛋白表达相关([r] < 1, P < 0.05)。结论长期暴露于环境浓度NP可引起大鼠免疫功能障碍,产生免疫毒性和炎症作用,并导致转录因子和差异免疫炎症因子活性的改变。图形抽象
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来源期刊
Environmental Sciences Europe
Environmental Sciences Europe Environmental Science-Pollution
CiteScore
9.20
自引率
1.70%
发文量
110
审稿时长
13 weeks
期刊介绍: ESEU is an international journal, focusing primarily on Europe, with a broad scope covering all aspects of environmental sciences, including the main topic regulation. ESEU will discuss the entanglement between environmental sciences and regulation because, in recent years, there have been misunderstandings and even disagreement between stakeholders in these two areas. ESEU will help to improve the comprehension of issues between environmental sciences and regulation. ESEU will be an outlet from the German-speaking (DACH) countries to Europe and an inlet from Europe to the DACH countries regarding environmental sciences and regulation. Moreover, ESEU will facilitate the exchange of ideas and interaction between Europe and the DACH countries regarding environmental regulatory issues. Although Europe is at the center of ESEU, the journal will not exclude the rest of the world, because regulatory issues pertaining to environmental sciences can be fully seen only from a global perspective.
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