Ding Hongyan, Yan Weizhe, Zhang Daoliang, Wang Lei, Yang Yue, Zhao Chang, Feng Shibin, Wang Xichun, Tang Jishun, Wu Dong, Wu Jinjie, Li Yu
College of Veterinary Medicine, Anhui Agricultural University, Hefei 230036, China.
Anhui Province Key Laboratory of Livestock and Poultry Product Safety Engineering, Institute of Animal Science and Veterinary Medicine, Anhui Academy of Agricultural Sciences, Hefei 230001, China.
Int J Mol Sci. 2025 May 16;26(10):4788. doi: 10.3390/ijms26104788.
Transition dairy cows face severe oxidative stress that disrupts mammary epithelial homeostasis through intertwined oxidative, inflammatory, and endoplasmic reticulum (ER) stress pathways. This study hypothesized that rutin, a natural flavonoid, alleviates hydrogen peroxide (HO)-induced oxidative damage in bovine mammary epithelial cells (BMECs) via AMPK/NFE2L2 signaling activation. In this study, BMECs were pre-incubated with rutin. Subsequently, cells were treated with or without HO. Additionally, by transfecting BMECs with NFE2L2 siRNA (siNFE2L2), we investigated how AMPK/NFE2L2 signaling mediated by rutin may prevent HO-induced oxidative damage. The results show that increases in reactive oxygen species (ROS), expression of inflammatory cytokines, expression of proteins related to endoplasmic reticulum stress and the apoptosis rate induced by HO in cells, were attenuated in rutin cultures. Challenges with HO led to a lower abundance of proteins related to AMPK and NFE2L2. Comparatively, these effects were reversed in cultures with rutin. Transfection with siNFE2L2 reversed the protection of rutin, suggesting that NFE2L2 is essential for the protective mechanism of rutin. These results elucidated the molecular mechanism of rutin's resistance to HO-mediated oxidative injury through the AMPK/NFE2L2 signaling pathway and suggested that it could be used as a potent in vivo antioxidant for ruminants during periods of stress, such as before and after calving.
围产期奶牛面临严重的氧化应激，这种应激通过相互交织的氧化、炎症和内质网（ER）应激途径破坏乳腺上皮细胞的稳态。本研究假设，天然黄酮类化合物芦丁通过激活AMPK/NFE2L2信号通路，减轻过氧化氢（H₂O₂）诱导的牛乳腺上皮细胞（BMECs）氧化损伤。在本研究中，BMECs先用芦丁预孵育。随后，细胞分别用或不用H₂O₂处理。此外，通过用NFE2L2 siRNA（siNFE2L2）转染BMECs，我们研究了芦丁介导的AMPK/NFE2L2信号通路如何预防H₂O₂诱导的氧化损伤。结果表明，芦丁培养的细胞中，活性氧（ROS）增加、炎症细胞因子表达、内质网应激相关蛋白表达以及H₂O₂诱导的细胞凋亡率均得到缓解。H₂O₂刺激导致与AMPK和NFE2L2相关的蛋白质丰度降低。相比之下，芦丁培养的细胞中这些效应得到逆转。用siNFE2L2转染可逆转芦丁的保护作用，表明NFE2L2对芦丁的保护机制至关重要。这些结果阐明了芦丁通过AMPK/NFE2L2信号通路抵抗H₂O₂介导的氧化损伤的分子机制，并表明它可作为反刍动物在应激期（如产犊前后）的一种有效的体内抗氧化剂。
