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（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。

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.