N-乙酰-半胱氨酸干预壬基酚对小鼠Sertoli TM4细胞的损伤作用

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(20μmol/L NP处理)、NP+NAC组(5 mmol/L NAC预处理4 h后20μmol/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,20μmol/L NP处理24 h能显著降低细胞存活率(P0.05),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。

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

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.