Shao Dan, Shen Wenxiang, Miao Yuyang, Gao Zhen, Pan Menghao, Wei Qiang, Yan Zuoting, Zhao Xiaoe, Ma Baohua
Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, College of Veterinary Medicine, Northwest A&F University, Yangling, 712100, Shaanxi, China.
Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Science, Lanzhou, 730050, China.
J Anim Sci Biotechnol. 2023 May 3;14(1):61. doi: 10.1186/s40104-023-00858-9.
Mastitis not only deteriorates the composition or quality of milk, but also damages the health and productivity of dairy goats. Sulforaphane (SFN) is a phytochemical isothiocyanate compound with various pharmacological effects such as anti-oxidant and anti-inflammatory. However, the effect of SFN on mastitis has yet to be elucidated. This study aimed to explore the anti-oxidant and anti-inflammatory effects and potential molecular mechanisms of SFN in lipopolysaccharide (LPS)-induced primary goat mammary epithelial cells (GMECs) and a mouse model of mastitis.
In vitro, SFN downregulated the mRNA expression of inflammatory factors (tumor necrosis factor-α (TNF-α), interleukin (IL)-1β and IL-6), inhibited the protein expression of inflammatory mediators (cyclooxygenase-2 (COX2), and inducible nitric oxide synthase (iNOS)) while suppressing nuclear factor kappa-B (NF-κB) activation in LPS-induced GMECs. Additionally, SFN exhibited an antioxidant effect by increasing Nrf2 expression and nuclear translocation, up-regulating antioxidant enzymes expression, and decreasing LPS-induced reactive oxygen species (ROS) production in GMECs. Furthermore, SFN pretreatment promoted the autophagy pathway, which was dependent on the increased Nrf2 level, and contributed significantly to the improved LPS-induced oxidative stress and inflammatory response. In vivo, SFN effectively alleviated histopathological lesions, suppressed the expression of inflammatory factors, enhanced immunohistochemistry staining of Nrf2, and amplified of LC3 puncta LPS-induced mastitis in mice. Mechanically, the in vitro and in vivo study showed that the anti-inflammatory and anti-oxidative stress effects of SFN were mediated by the Nrf2-mediated autophagy pathway in GMECs and a mouse model of mastitis.
These results indicate that the natural compound SFN has a preventive effect on LPS-induced inflammation through by regulating the Nrf2-mediated autophagy pathway in primary goat mammary epithelial cells and a mouse model of mastitis, which may improve prevention strategies for mastitis in dairy goats.
乳腺炎不仅会降低羊奶的成分或质量，还会损害奶山羊的健康和生产力。萝卜硫素（SFN）是一种具有多种药理作用（如抗氧化和抗炎）的植物化学异硫氰酸酯化合物。然而，SFN对乳腺炎的影响尚未阐明。本研究旨在探讨SFN在脂多糖（LPS）诱导的原代山羊乳腺上皮细胞（GMECs）和乳腺炎小鼠模型中的抗氧化和抗炎作用及潜在分子机制。
在体外，SFN下调炎症因子（肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-1β和IL-6）的mRNA表达，抑制炎症介质（环氧化酶-2（COX2）和诱导型一氧化氮合酶（iNOS））的蛋白表达，同时抑制LPS诱导的GMECs中核因子κB（NF-κB）的激活。此外，SFN通过增加Nrf2表达和核转位、上调抗氧化酶表达以及减少LPS诱导的GMECs中活性氧（ROS）的产生而表现出抗氧化作用。此外，SFN预处理促进了自噬途径，这依赖于Nrf2水平的升高，并显著改善了LPS诱导的氧化应激和炎症反应。在体内，SFN有效减轻组织病理学损伤，抑制炎症因子的表达，增强Nrf2的免疫组化染色，并增加LPS诱导的小鼠乳腺炎中LC3斑点。从机制上讲，体外和体内研究表明，SFN的抗炎和抗氧化应激作用是由GMECs和乳腺炎小鼠模型中Nrf2介导的自噬途径介导的。
这些结果表明，天然化合物SFN通过调节原代山羊乳腺上皮细胞和乳腺炎小鼠模型中Nrf2介导的自噬途径，对LPS诱导的炎症具有预防作用，这可能改善奶山羊乳腺炎的预防策略。
