壬基酚异构体NP42对小鼠RAW264.7巨噬细胞的损伤作用
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(1.东莞理工学院化学工程与能源技术学院 食品营养健康与智能化加工研究中心 广东东莞 523808;2.南昌大学 食品科学与技术国家重点实验室 南昌 330047)

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刘晓珍(1990—),女,博士,副教授

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基金项目:

国家自然科学基金青年科学基金项目(81803193);广东省自然科学基金项目(2018A030310033)


The Damage Effect of Nonylphenol Isomer NP42 on Mouse Macrophage RAW264.7 Cells
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(1.Engineering Research Center of Health Food Design & Nutrition Regulation, School of Chemical Engineering and Energy Technology, Dongguan University of Technology, Dongguan 523808, Guangdong;2.State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047)

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    摘要:

    目的:研究壬基酚异构体NP42对小鼠巨噬细胞RAW264.7的损伤作用并探究其分子机制。方法:以小鼠巨噬细胞RAW264.7为研究对象,将不同浓度NP42(0.1~100 μmol/L)作用于细胞24 h,采用噻唑蓝法检测细胞存活率,酶联免疫分析检测环磷酸鸟苷(cGMP)和环磷酸腺苷(cAMP)含量,RT-PCR法检测肿瘤坏死因子α(TNF-α)和诱导型一氧化氮合成酶(iNOS)mRNA的表达水平,Western blot法检测蛋白激酶C(PKC)的表达情况以及p38丝裂原活化蛋白激酶(p38-MAPK)的磷酸化水平。结果:0.1~10 μmol/L NP42对细胞存活率无显著影响,100 μmol/L NP42能显著降低RAW264.7细胞的存活率(P < 0.01)。与对照组相比,NP42处理24 h能显著抑制细胞TNF-α mRNA和iNOS mRNA的表达,抑制细胞PKC蛋白的表达和降低细胞内cAMP的含量,同时增强细胞内cGMP的含量。NP42处理能显著下调p38的磷酸化水平。结论:壬基酚异构体NP42通过PKC-cAMP信号通路以及p38-MAPK信号通路对小鼠巨噬细胞RAW264.7产生损伤作用,这可能是壬基酚发挥免疫损伤的主要分子机制。

    Abstract:

    Objective: The aim of this study was to investigate the damage effect of nonylphenol isomer NP42 on mouse macrophage RAW264.7 cells and the underlying molecular mechanism. Methods: The RAW264.7 cells were with various concentration of NP42 for 24 h, then the cell survival rate was detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay, the cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP) contents were detected by commercial kits, RT-PCR was used to detect the mRNA expression level of tumor necrosis factor-α (TNF-α) and inducible nitric oxides synthase (iNOS), Western blot was used to detect the phosphorylation level of p38 mitogen-activated protein kinase (p38 MAPK) and the expression of protein kinase C (PKC). Results: Compared with the Control group, 0.1-10 μmol/L NP42 showed no effect on cell survival, while NP42 extremely decreased cell survival at 100 μmol/L; Moreover, NP42 treatment significantly inhibited the mRNA expression of TNF-α and iNOS, the expression of PKC protein and the content of cAMP in cells. In contrast, NP42 upregulate the content of cGMP in the cells. In addition, NP42 inhibited the phosphorylation level of p38-MAPK. Conclusion: PKC-cAMP pathway and p38 MAPK signaling pathway were involved in nonylphenol isomer NP42 induced damage in RAW264.7 cells, and this should be one of the major mechanisms by which nonylphenol exerted immunotoxic effects in organism.

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刘晓珍,卢培泉,李福香,祝兆亮,苏梓烁,黄丹菲,谢明勇.壬基酚异构体NP42对小鼠RAW264.7巨噬细胞的损伤作用[J].中国食品学报,2022,22(10):58-65

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  • 收稿日期:2021-10-19
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  • 在线发布日期: 2022-11-24
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