Laboratory of food Chemistry and Toxicology, Faculty of Pharmacy, University of Valencia, Av. Vicent Andrés Estellés s/n, 46100, Burjassot, València, Spain.
Laboratory of food Chemistry and Toxicology, Faculty of Pharmacy, University of Valencia, Av. Vicent Andrés Estellés s/n, 46100, Burjassot, València, Spain.
Toxicol In Vitro. 2023 Dec;93:105670. doi: 10.1016/j.tiv.2023.105670. Epub 2023 Aug 24.
The presence of mycotoxins such as Fumonisin B1(FB1) and Ochratoxin A (OTA) in food and feed has become a threat to human and animal health since they can produce several afflictions. Different mechanisms of action by which they exercise their cytotoxic activity have been attributed to them, including the production of reactive oxygen species (ROS). For this reason, a measurement of the production of ROS species, and an evaluation of the intrinsic cell enzymatic antioxidant activity, including glutathione peroxidase (GPx), glutathione transferase (GTS), and catalase (CAT) together with a cytotoxicity and cell cycle assay have been performed in undifferentiated SH-SY5Y cells exposed to FB1, OTA and [FB1 + OTA] after 24 h and 48 h. FB1 and OTA. Monitoring of intracellular ROS production was carried out by the H-DCFDA probe; while spectrometry analysis of absorbances was used for measuring GPx, GST and CAT activity. Finally, cell proliferation and cell cycle distribution were studied by flow cytometry. When cells were treated with OTA, an increase in GPx and GST activity was observed compared to FB1 and [FB1 + OTA]; conversely, a decrease in CAT activity was observed when cells were exposed to OTA coinciding with the results observed for ROS measurement. Regarding the cell cycle, when cells were exposed to OTA, a decrease in G0/G1 was detected, revealing an arrest of cell division for SH-SY5Y cells at the concentrations studied.
真菌毒素如伏马菌素 B1（FB1）和赭曲霉毒素 A（OTA）在食品和饲料中的存在对人类和动物健康构成了威胁，因为它们会产生多种疾病。它们被归因于几种不同的细胞毒性作用机制，包括活性氧（ROS）的产生。出于这个原因，在未分化的 SH-SY5Y 细胞中进行了 ROS 物种产生的测量，并评估了内在细胞酶抗氧化活性，包括谷胱甘肽过氧化物酶（GPx）、谷胱甘肽转移酶（GTS）和过氧化氢酶（CAT），以及细胞毒性和细胞周期测定，这些细胞在暴露于 FB1、OTA 和 [FB1+OTA] 24 小时和 48 小时后。FB1 和 OTA。通过 H-DCFDA 探针监测细胞内 ROS 的产生；同时通过光谱分析吸光度来测量 GPx、GST 和 CAT 活性。最后，通过流式细胞术研究细胞增殖和细胞周期分布。当细胞用 OTA 处理时，与 FB1 和 [FB1+OTA] 相比，观察到 GPx 和 GST 活性增加；相反，当细胞暴露于 OTA 时，CAT 活性下降，与 ROS 测量结果一致。关于细胞周期，当细胞暴露于 OTA 时，检测到 G0/G1 减少，这表明 SH-SY5Y 细胞在研究浓度下的细胞分裂停滞。
