Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection & Internet Technology, Zhejiang International Science and Technology Cooperation Base for Veterinary Medicine and Health Management, China-Australia Joint Laboratory for Animal Health Big Data Analytics, College of Animal Science and Technology & College of Veterinary Medicine of Zhejiang A&F University, Hangzhou, 311300, People's Republic of China.
Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection & Internet Technology, Zhejiang International Science and Technology Cooperation Base for Veterinary Medicine and Health Management, China-Australia Joint Laboratory for Animal Health Big Data Analytics, College of Animal Science and Technology & College of Veterinary Medicine of Zhejiang A&F University, Hangzhou, 311300, People's Republic of China.
Theriogenology. 2024 Jan 15;214:192-200. doi: 10.1016/j.theriogenology.2023.10.028. Epub 2023 Oct 25.
Fluorochloridone (FLC), a pyrrolidone herbicide, has been recognized as a hazardous chemical. The in vitro adverse effects of FLC on the reproduction of livestock have not been assessed. This study was conducted to explore the cytotoxicity and toxicological mechanisms of FLC on cultured goat Sertoli cells. The results showed that FLC exposure significantly decreased goat Sertoli cell viability (p < 0.05) and induced oxidative stress. And FLC treatment promoted apoptosis and initiation of autophagy. Interestingly, FLC inhibited lysosomal biogenesis and blocked autophagic flux in goat Sertoli cells. The expression levels of autophagy-related proteins Atg5, LC3II, and p62 were significantly increased (p < 0.05) in FLC-treated goat Sertoli cells compared with the control. Importantly, FLC-induced ROS accumulation further causes mitochondrial dysfunction and disturbs mitophagy. FLC significantly decreased (p < 0.05) the expression levels of OPA1, MFN2, p-Drp1, FIS1, PINK1, and Parkin in goat Sertoli cells. Moreover, pretreatment with N-acetyl-l-cysteine (NAC, an antioxidant) significantly reduced (p < 0.01) FLC-induced ROS accumulation and reversed the disorder of autophagy levels. Our results indicated that FLC-induced toxicity in primary goat Sertoli cells was characterized by ROS accumulation, inducing oxidative stress, inhibiting lysosomal biogenesis, blocking autophagic flux, and promoting mitochondrial dysfunction, resulting in apoptosis via the mitochondrial pathway.
氟氯草定（FLC）是一种吡咯烷类除草剂，已被认为是一种危险化学品。尚未评估 FLC 对家畜繁殖的体外不良影响。本研究旨在探讨 FLC 对培养的山羊支持细胞的细胞毒性和毒理学机制。结果表明，FLC 暴露显著降低了山羊支持细胞的活力（p<0.05）并诱导了氧化应激。FLC 处理促进了细胞凋亡和自噬的起始。有趣的是，FLC 抑制了山羊支持细胞中的溶酶体生物发生并阻断了自噬流。与对照组相比，FLC 处理的山羊支持细胞中自噬相关蛋白 Atg5、LC3II 和 p62 的表达水平显著增加（p<0.05）。重要的是，FLC 诱导的 ROS 积累进一步导致线粒体功能障碍并扰乱了线粒体自噬。FLC 显著降低了（p<0.05）山羊支持细胞中 OPA1、MFN2、p-Drp1、FIS1、PINK1 和 Parkin 的表达水平。此外，用 N-乙酰-L-半胱氨酸（NAC，一种抗氧化剂）预处理可显著减少（p<0.01）FLC 诱导的 ROS 积累并逆转自噬水平的紊乱。我们的结果表明，FLC 诱导的原代山羊支持细胞毒性的特征是 ROS 积累，诱导氧化应激，抑制溶酶体生物发生，阻断自噬流，并通过线粒体途径导致细胞凋亡和线粒体功能障碍。
