Data Sheet 1_Evaluation the protective role of baicalin against H2O2-driven oxidation, inflammation and apoptosis in bovine mammary epithelial cells.xlsx

Mastitis is one of the most common diseases in dairy farms. During the perinatal period, the bovine mammary epithelial cells (BMECs) of High-yielding dairy cows accelerate metabolism and produce large amounts of reactive oxygen species (ROS). It is one of the primary causes of mastitis and will lead to the breakdown of redox balance, which will induce oxidative stress, inflammation, and apoptosis. Baicalin is a flavonoid substance extracted from the root of natural plant Scutellaria baicalensis, which has anti-inflammatory, anti-oxidant, anti-viral and other biological functions. In this research, hydrogen peroxide (H2O2) was used to construct a mastitis oxidative stress model, and relevant mechanisms were analyzed by immunofluorescence techniques, qRT-PCR and Western Blot to explore how baicalin affects BMECs' oxidative stress and inflammation caused by H2O2, as well as to provide new perspectives on the combined application of baicalin in the prevention and treatment of mastitis. The results demonstrated that baicalin treatment could reduce the accumulation of H2O2-induced intracellular ROS and decrease the expression of inflammatory cytokines Tumor Necrosis Factor-α (TNF-α), interleukin 6 (IL-6), interleukin-1β (IL-1β) and the apoptosis rate. The inhibitory effect of baicalin on H2O2-induced intracellular ROS accumulation and the expression of inflammatory cytokines and apoptotic factors in BMECs was blocked by pretreatment with the Nuclear factor erythroid 2-related factor 2 (Nrf2) inhibitor retinoic acid (RA) prior to H2O2 and/or baicalin treatment. In summary, baicalin could served as a natural antioxidant agent to regulate cell apoptosis through its anti-inflammatory, antioxidant and anti-apoptotic effects to combat BMECs damage caused by H2O2.

乳腺炎（Mastitis）是奶牛养殖场最常见的疾病之一。围产期高产奶牛的乳腺上皮细胞（bovine mammary epithelial cells, BMECs）代谢速率显著加快，并会产生大量活性氧（reactive oxygen species, ROS），这是引发乳腺炎的核心诱因之一；该过程会破坏机体氧化还原平衡，进而诱导氧化应激、炎症反应与细胞凋亡。黄芩苷（baicalin）是从天然植物黄芩（Scutellaria baicalensis）根中提取的黄酮类物质，具备抗炎、抗氧化、抗病毒等多种生物学功能。本研究采用过氧化氢（hydrogen peroxide, H₂O₂）构建乳腺炎氧化应激细胞模型，通过免疫荧光技术、qRT-PCR与蛋白质印迹（Western Blot）分析相关作用机制，旨在探究黄芩苷对过氧化氢诱导的奶牛乳腺上皮细胞氧化应激与炎症反应的影响，为黄芩苷在乳腺炎防治领域的联合应用提供全新研究视角。实验结果表明，黄芩苷处理可有效减少过氧化氢诱导的细胞内活性氧积累，降低促炎细胞因子肿瘤坏死因子-α（Tumor Necrosis Factor-α, TNF-α）、白细胞介素6（interleukin 6, IL-6）、白细胞介素-1β（interleukin-1β, IL-1β）的表达水平，并抑制细胞凋亡率。若在过氧化氢和/或黄芩苷处理前，使用核因子E2相关因子2（Nuclear factor erythroid 2-related factor 2, Nrf2）抑制剂视黄酸（retinoic acid, RA）进行预处理，则可阻断黄芩苷对过氧化氢诱导的奶牛乳腺上皮细胞内活性氧积累、促炎细胞因子及凋亡因子表达的抑制作用。综上，黄芩苷可作为天然抗氧化剂，通过其抗炎、抗氧化及抗凋亡活性调控细胞凋亡，以对抗过氧化氢诱导的奶牛乳腺上皮细胞损伤。