College of Veterinary Medicine, Yangzhou University, 12 Wenhui East Road, Yangzhou 225009, China.
Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China.
Toxins (Basel). 2022 Oct 26;14(11):733. doi: 10.3390/toxins14110733.
Zearalenone (ZEA) is a common mycotoxin that induces oxidative stress (OS) and affects the male reproductive system in animals. Resveratrol (RSV) has good antioxidant activity and can activate nuclear factor erythroid 2-related factor (Nrf2) to protect cells through the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway. The objective of this study was to investigate the protective effect and the mechanism of RSV on OS and apoptosis in TM4 cells induced by ZEA. Prior to being exposed to ZEA, TM4 cells were pretreated with RSV or the PI3K/Akt inhibitor LY294002. Cell viability was measured by Cell Counting Kit-8 (CCK-8) assays. Flow cytometry was used to determine the level of apoptosis and intracellular reactive oxygen species (ROS). The expression of poly ADP-ribose polymerase (PARP), caspase-3, BCL2-associated X (Bax)/B-cell lymphoma-2 (Bcl-2), and PI3K/Akt-mediated Nrf2/heme oxygenase 1 (HO-1) signaling pathway-related proteins was evaluated by Western blotting. Nrf2 siRNA transfection and LY294002 treatment were used to investigate the role of the Nrf2/HO-1 and PI3K/Akt signaling pathways in RSV alleviation of ZEA-induced OS. The results showed that pretreatment with RSV significantly reduced the expression of apoptosis-related proteins and increased cell viability. Catalase (CAT) activity and glutathione (GSH) levels were also increased, whereas malondialdehyde (MDA) and ROS levels decreased ( < 0.05). RSV also upregulated Akt phosphorylation, Nrf2 nuclear translocation, and HO-1 expression under conditions of OS ( < 0.05). Transfection with Nrf2 siRNA abolished the protective effects of RSV against ZEA-induced cytotoxicity ( < 0.05), ROS accumulation ( < 0.05), and apoptosis ( < 0.05). LY294002 completely blocked the RSV-mediated increase in Nrf2 nuclear translocation ( < 0.05), HO-1 expression ( < 0.05), and cytoprotective activity ( < 0.05). Collectively, the above findings indicate that RSV can protect against ZEA-induced OS and apoptosis in TM4 cells by PI3K/Akt-mediated activation of the Nrf2/HO-1 signaling pathway.
玉米赤霉烯酮（ZEA）是一种常见的真菌毒素，可诱导氧化应激（OS）并影响动物的雄性生殖系统。白藜芦醇（RSV）具有良好的抗氧化活性，可通过磷脂酰肌醇 3-激酶（PI3K）/蛋白激酶 B（Akt）信号通路激活核因子红细胞 2 相关因子（Nrf2）来保护细胞。本研究旨在探讨 RSV 对 ZEA 诱导的 TM4 细胞 OS 和凋亡的保护作用及其机制。在暴露于 ZEA 之前，用 RSV 或 PI3K/Akt 抑制剂 LY294002 预处理 TM4 细胞。通过细胞计数试剂盒-8（CCK-8）测定法测量细胞活力。通过流式细胞术测定细胞凋亡和细胞内活性氧（ROS）水平。通过 Western blot 评估多聚 ADP-核糖聚合酶（PARP）、半胱天冬氨酸蛋白酶-3（caspase-3）、B 细胞淋巴瘤-2 相关 X（Bax）/B 细胞淋巴瘤-2（Bcl-2）以及 PI3K/Akt 介导的 Nrf2/血红素加氧酶 1（HO-1）信号通路相关蛋白的表达。用 Nrf2 siRNA 转染和 LY294002 处理来研究 Nrf2/HO-1 和 PI3K/Akt 信号通路在 RSV 减轻 ZEA 诱导的 OS 中的作用。结果表明，RSV 预处理可显著降低凋亡相关蛋白的表达并增加细胞活力。过氧化氢酶（CAT）活性和谷胱甘肽（GSH）水平也增加，而丙二醛（MDA）和 ROS 水平降低（<0.05）。RSV 还可在 OS 条件下上调 Akt 磷酸化、Nrf2 核易位和 HO-1 表达（<0.05）。用 Nrf2 siRNA 转染可消除 RSV 对 ZEA 诱导的细胞毒性（<0.05）、ROS 积累（<0.05）和凋亡（<0.05）的保护作用。LY294002 完全阻断 RSV 介导的 Nrf2 核易位（<0.05）、HO-1 表达（<0.05）和细胞保护活性（<0.05）的增加。综上所述，这些发现表明 RSV 可通过 PI3K/Akt 介导的 Nrf2/HO-1 信号通路的激活来保护 TM4 细胞免受 ZEA 诱导的 OS 和凋亡。
