食品科学 ›› 2019, Vol. 40 ›› Issue (13): 130-136.doi: 10.7506/spkx1002-6630-20180614-242

• 营养卫生 • 上一篇    下一篇

N-乙酰-半胱氨酸干预壬基酚对小鼠Sertoli TM4细胞的损伤作用

刘晓珍,聂少平,余 强,黄丹菲,谢明勇   

  1. 1.东莞理工学院化学工程与能源技术学院,科技创新研究院,广东 东莞 523808;2.南昌大学 食品科学与技术国家重点实验室,江西 南昌 330047
  • 出版日期:2019-07-15 发布日期:2019-07-23
  • 基金资助:
    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);东莞理工学院科研启动专项经费项目(GC300502-33);东莞市一般社科项目(20185071401508)

N-Acetyl-cysteine Attenuates Nonylphenol-Induced Damage in Mouse Sertoli TM4 Cells

LIU Xiaozhen, NIE Shaoping, YU Qiang, HUANG Danfei, XIE Mingyong   

  1. 1. Institute of Science and Technology Innovation, School of Chemical Engineering and Energy Technology, Dongguan University of Technology, Dongguan 523808, China; 2. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China
  • Online:2019-07-15 Published:2019-07-23

摘要: 目的:研究N-乙酰-半胱氨酸(N-acetyl-cysteine,NAC)对壬基酚(nonylphenol,NP)诱导的小鼠Sertoli TM4细胞氧化损伤及凋亡的干预作用。方法:以Sertoli TM4细胞为对象,实验分为对照组、NP组(20 μmol/L NP处理)、NP+NAC组(5 mmol/L NAC预处理4 h后20 μmol/L NP处理24 h)、NAC组(5 mmol/L NAC处理4 h后换正常培养基培养),采用噻唑蓝法检测细胞存活率;流式细胞术检测活性氧(reactive oxygen species,ROS)含量和细胞凋亡情况;试剂盒法检测超氧化物歧化酶(superoxide dismutase,SOD)活力、过氧化氢酶(catalase,CAT)活力、丙二醛(malondialdehyde,MDA)含量及Caspase-3相对活力;Western blot法检测细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)和c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)磷酸化情况。结果:与对照组相比,20 μmol/L NP处理24 h能显著降低细胞存活率(P<0.05),同时诱导细胞内ROS生成,下调SOD和CAT活力,增加MDA含量,诱导Sertoli TM4细胞凋亡,增加Caspase-3相对活力,促进ERK、JNK蛋白磷酸化激活;与NP组相比,NAC预处理能够明显削弱NP引起的细胞内ROS生成,使SOD、CAT活力下调,MDA含量增加,Sertoli TM4细胞凋亡,Caspase-3相对活力增强,激活ERK、JNK信号通路。结论:NAC具有干预NP对小鼠Sertoli TM4细胞损伤的作用,这可能与NAC抑制NP诱导的细胞氧化应激和凋亡以及阻断ERK、JNK信号通路的激活相关。

关键词: 壬基酚, 小鼠Sertoli TM4细胞, N-乙酰-半胱氨酸, 细胞凋亡, 氧化应激

Abstract: Objective: The aim of this study was to explore the effect of N-acetyl-cysteine (NAC) on nonylphenol (NP)-induced oxidative stress and cell apoptosis in TM4 mouse Sertoli cells. Methods: The cells were divided into 4 groups: control group, NP group (20 μmol/L NP), NP + NAC group (sequential treatment with 5 mmol/L NAC for 4 h followed by 20 mmol/L NP for 24 h), NAC group (sequential treatment with 5 mmol/L NAC for 4 h followed by normal cell culture medium). Cell survival rate was monitored by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay, and reactive oxygen species (ROS) generation and cell apoptosis were analyzed by flow cytometry. Caspase-3 activity, superoxide dismutase (SOD) activity, catalase (CAT) activity, and malondialdehyde (MDA) content were measured by assay kits. The phosphorylation of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) was detected by Western blot assay. Results: Compared with the control group, NP decreased cell survival rate significantly (P < 0.05), induced reactive oxygen species (ROS) generation, increased SOD and CAT activities, and decreased MDA content in Sertoli TM4 cells. Additionally, NP induced cell apoptosis, increased caspase-3 activity, and activated the ERK/JNK signaling pathways. Compared with the NP group, NAC pretreatment attenuated ROS production, reduced SOD and CAT activities and increased MDA content. Moreover, NAC blocked cell apoptosis, increased caspase-3 activity, and activated the ERK/JNK signal pathways. Conclusion: NAC attenuated NP-induced damage in mouse Sertoli TM4 cells, which may be related to the ability of NAC to attenuate NP-induced oxidative stress and cell apoptosis and block the activation of the ERK and JNK signaling pathways.

Key words: nonylphenol, mouse Sertoli TM4 cells, N-acetyl-cysteine, cell apoptosis, oxidative stress

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