Toxicoproteomic study of fipronil in SH-SY5Y cells reveals induction of endoplasmic reticulum stress and necrotic cell death as neurodegenerative mechanisms

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro Pub Date : 2025-05-29 DOI:10.1016/j.tiv.2025.106098

Theetat Ruangjaroon , N. Monique Paricharttanakul , Daranee Chokchaichamnankit , Chantragan Srisomsap , Kriengsak Lirdprapamongkol , Jisnuson Svasti

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Abstract

Exposure to pesticides has been considered as a risk factor for developing neurodegenerative diseases. The increasing use of fipronil, a phenylpyrazole insecticide, poses a risk to human health. This study aims to use toxicoproteomics for exploring neurodegenerative mechanism of fipronil in SH-SY5Y human neuroblastoma cells. In this study, fipronil at sub-cytotoxic and cytotoxic concentrations (43 and 78 μM) caused increases in superoxide level from 3 to 48 h after treatment, while intracellular glutathione level was decreased at 48 h. Neurite outgrowth of the cells was impaired by fipronil at both concentrations, while significant increase of cell death via apoptosis and necrosis modes were observed with fipronil at cytotoxic concentration. Pretreatment with antioxidant N-acetylcysteine (NAC) effectively relieved impairment of neurite outgrowth and induction of cell death by fipronil. Proteomic analysis showed that expression of proteins involving endoplasmic reticulum (ER) stress and unfolded protein responses were predominantly affected by fipronil. Immunoblotting confirmed the increased expression of ER stress markers, GRP78/BiP (78 kDa glucose-regulated protein/Binding immunoglobulin protein) and PDI (protein disulfide isomerase), in fipronil-treated cells. Improved understanding of the neurotoxic mechanism of fipronil may help in developing a strategy for reducing risk of neurodegenerative development from intense and prolonged use of fipronil.

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氟虫腈在SH-SY5Y细胞中的毒性蛋白质组学研究揭示了内质网应激和坏死细胞死亡是神经退行性机制

接触杀虫剂被认为是发生神经退行性疾病的一个危险因素。越来越多地使用苯基吡唑类杀虫剂氟虫腈，对人类健康构成威胁。本研究旨在利用毒理学蛋白质组学方法探讨氟虫腈对SH-SY5Y人神经母细胞瘤细胞的神经退行性作用机制。在本研究中，亚细胞毒性和细胞毒性浓度（43 μM和78 μM）的氟虫腈在处理后3 ~ 48 h内引起超氧化物水平升高，而细胞内谷胱甘肽水平在48 h内降低。两种浓度的氟虫腈均损害细胞的神经突生长，同时细胞毒性浓度的氟虫腈显著增加细胞凋亡和坏死模式的死亡。抗氧化剂n -乙酰半胱氨酸（NAC）预处理可有效缓解氟虫腈诱导的神经突生长损伤和细胞死亡。蛋白质组学分析显示，参与内质网应激和未折叠蛋白反应的蛋白质表达主要受到氟虫腈的影响。免疫印迹证实，在氟虫腈处理的细胞中，内质网应激标志物GRP78/BiP （78 kDa葡萄糖调节蛋白/结合免疫球蛋白蛋白）和PDI（蛋白二硫异构酶）的表达增加。提高对氟虫腈神经毒性机制的了解可能有助于制定一种策略，以减少因长期高强度使用氟虫腈而导致的神经退行性发展的风险。

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来源期刊

Toxicology in Vitro 医学-毒理学

CiteScore

6.50

自引率

3.10%

发文量

181

审稿时长

65 days

期刊介绍： Toxicology in Vitro publishes original research papers and reviews on the application and use of in vitro systems for assessing or predicting the toxic effects of chemicals and elucidating their mechanisms of action. These in vitro techniques include utilizing cell or tissue cultures, isolated cells, tissue slices, subcellular fractions, transgenic cell cultures, and cells from transgenic organisms, as well as in silico modelling. The Journal will focus on investigations that involve the development and validation of new in vitro methods, e.g. for prediction of toxic effects based on traditional and in silico modelling; on the use of methods in high-throughput toxicology and pharmacology; elucidation of mechanisms of toxic action; the application of genomics, transcriptomics and proteomics in toxicology, as well as on comparative studies that characterise the relationship between in vitro and in vivo findings. The Journal strongly encourages the submission of manuscripts that focus on the development of in vitro methods, their practical applications and regulatory use (e.g. in the areas of food components cosmetics, pharmaceuticals, pesticides, and industrial chemicals). Toxicology in Vitro discourages papers that record reporting on toxicological effects from materials, such as plant extracts or herbal medicines, that have not been chemically characterized.