壬基酚异构体NP_(42)对小鼠RAW264.7巨噬细胞的损伤作用  

The Damage Effect of Nonylphenol Isomer NP_(42) on Mouse Macrophage RAW264.7 Cells

作　　者：刘晓珍 卢培泉[1] 李福香 祝兆亮 苏梓烁 黄丹菲 谢明勇[2] Liu Xiaozhen;Lu Peiquan;Li Fuxiang;Zhu Zhaoliang;Su Zishuo;Huang Danfei;Xie Mingyong(Engineering Research Center of Health Food Design&Nutrition Regulation,School of Chemical Engineering and Energy Technology,Dongguan University of Technology,Dongguan 523808,Guangdong;State Key Laboratory of Food Science and Technology,Nanchang University,Nanchang 330047)

机构地区：[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

出　　处：《中国食品学报》2022年第10期58-65,共8页Journal of Chinese Institute Of Food Science and Technology

基　　金：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)。

摘　　要：目的:研究壬基酚异构体NP_(42)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NP_(42)(0.1~100μmol/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:0.1~10μmol/L NP对细胞存活率无显著影响,100μmol/L NP_(42)能显著降低RAW264.7细胞的存活率(P <0.01)。与对照组相比,NP_(42)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c GMP的含量。NP_(42)处理能显著下调p38的磷酸化水平。结论:壬基酚异构体NP_(42)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。Objective: The aim of this study was to investigate the damage effect of nonylphenol isomer NP_(42)on mouse macrophage RAW264.7 cells and the underlying molecular mechanism. Methods: The RAW264.7 cells were with various concentration of NP_(42) 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 NP_(42) showed no effect on cell survival, while NP_(42) extremely decreased cell survival at 100 μmol/L;Moreover, NP_(42) 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, NP_(42) upregulate the content of cGMP in the cells. In addition, NP_(42) inhibited the phosphorylation level of p38-MAPK. Conclusion: PKC-cAMP pathway and p38 MAPK signaling pathway were involved in nonylphenol isomer NP_(42) induced damage in RAW264.7 cells, and this should be one of the major mechanisms by which nonylphenol exerted immunotoxic effects in organism.

关 键 词：壬基酚异构体NP_(42) 巨噬细胞RAW264.7 环磷酸腺苷 蛋白激酶C p38-MAPK信号通路 

分 类 号：TS201.4[轻工技术与工程—食品科学]