Lu Yujiao, Luo Qin, Cui Hengmin, Deng Huidan, Kuang Ping, Liu Huan, Fang Jing, Zuo Zhicai, Deng Junliang, Li Yinglun, Wang Xun, Zhao Ling
College of Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu 611130, China.
Key Laboratory of Animal Diseases and Environmental Hazards of Sichuan Province, Sichuan Agriculture University, Wenjiang, Chengdu 611130, China.
Aging (Albany NY). 2017 Jun 27;9(6):1623-1639. doi: 10.18632/aging.101257.
The current study was conducted to investigate the effect of sodium fluoride (NaF) on the oxidative stress and apoptosis as well as their relationship in the mouse liver by using methods of flow cytometry, quantitative real-time polymerase chain reaction (qRT-PCR), western blot, biochemistry and experimental pathology. 240 four-week-old ICR mice were randomly divided into 4 groups and exposed to different concentration of NaF (0 mg/kg, 12 mg/kg, 24 mg/kg and 48 mg/kg) for a period of 42 days. The results showed that NaF caused oxidative stress and apoptosis. NaF-caused oxidative stress was accompanied by increasing reactive oxygen species (ROS) and malondialdehyde (MDA) levels, and decreasing mRNA expression levels and activities of superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), glutathione peroxidase (GSH-PX) and glutathione-s-transferase (GST). NaF induced apoptosis via tumor necrosis factor recpter-1 (TNF-R1) signaling pathway, which was characterized by significantly increasing mRNA and protein expression levels of TNF-R1, Fas associated death domain (FADD), TNFR-associated death domain (TRADD), cysteine aspartate specific protease-8 (caspase-8) and cysteine aspartate specific protease-3 (caspase-3) in dose- and time-dependent manner. Oxidative stress is involved in the process of apoptotic occurrence, and can be triggered by promoting ROS production and reducing antioxidant function. NaF-caused oxidative stress and apoptosis finally impaired hepatic function, which was strongly supported by the histopathological lesions and increased serum alanine amino transferase (ALT), aspartic acid transferase (AST), alkaline phosphatase (AKP) activities and TBIL contents.
本研究旨在通过流式细胞术、定量实时聚合酶链反应（qRT-PCR）、蛋白质免疫印迹法、生物化学和实验病理学方法，探讨氟化钠（NaF）对小鼠肝脏氧化应激和细胞凋亡的影响及其相互关系。将240只4周龄的ICR小鼠随机分为4组，分别暴露于不同浓度的NaF（0 mg/kg、12 mg/kg、24 mg/kg和48 mg/kg）中42天。结果表明，NaF可引起氧化应激和细胞凋亡。NaF引起的氧化应激表现为活性氧（ROS）和丙二醛（MDA）水平升高，超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽（GSH）、谷胱甘肽过氧化物酶（GSH-PX）和谷胱甘肽-S-转移酶（GST）的mRNA表达水平和活性降低。NaF通过肿瘤坏死因子受体-1（TNF-R1）信号通路诱导细胞凋亡，其特征是TNF-R1、Fas相关死亡结构域（FADD）、TNFR相关死亡结构域（TRADD）、半胱氨酸天冬氨酸特异性蛋白酶-8（caspase-8）和半胱氨酸天冬氨酸特异性蛋白酶-3（caspase-3）的mRNA和蛋白质表达水平呈剂量和时间依赖性显著增加。氧化应激参与细胞凋亡的发生过程，可通过促进ROS产生和降低抗氧化功能而引发。NaF引起的氧化应激和细胞凋亡最终损害肝功能，组织病理学损伤以及血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、碱性磷酸酶（AKP）活性和总胆红素（TBIL）含量升高有力地支持了这一点。
