樱黄素通过Nrf2通路抑制脂多糖诱导的奶牛乳腺上皮细胞炎症反应和脂质过氧化

doi:10.3969/j.issn.1674-7968.2024.07.008

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摘要乳房炎对奶牛(Bos taurus)的健康和产奶量产生极大的影响,导致乳制品质量降低,给整个奶牛产业带来了巨大的经济损失。樱黄素(prunetin, Pru)具有抗过敏、抗炎等生物活性。本研究旨在探索樱黄素对脂多糖诱导的奶牛乳腺上皮细胞(bovine mammary epithelial cells, BMECs)炎症反应和脂质过氧化的影响及其机制。将实验分为对照(CK)组、脂多糖(lipopolysaccharide, LPS)(10 μg/mL)组、脂多糖添加樱黄素(10, 20, 40和80 μmol/L)(LPS+Pru)组和脂多糖加铁死亡抑制剂(ferrostatin-1, Fer-1)(10 μmol/L)(LPS+Fer-1)组,分别检测炎症因子、核因子κB (nuclear factor κB, NF-κB)和核转录相关因子(nuclear factor erythroid 2-related factor 2, Nrf2)/Kelch样ECH关联蛋白1 (Kelch-like ECH-associated protein 1, Keap1)信号通路蛋白的表达水平。结果显示,LPS可引起肿瘤坏死因子-α (tumor necrosis factor-α, TNF-α)、白细胞介素-1β (interleukin-1β, IL-1β)、IL-6和IL-8等炎症因子的基因和蛋白水平显著上调。LPS提高Fe²⁺、活性氧(reactive oxygen species, ROS)和丙二醛(malondialdehyde, MDA)含量。樱黄素可下调TNF-α、IL-1β、IL-6和IL-8基因及其蛋白的表达,上调Nrf2基因及其蛋白表达水平,抑制NF-κB信号通路,降低细胞死亡率。樱黄素还可降低Fe²⁺、ROS和MDA含量,提高谷胱甘肽过氧化物酶4 (glutathione peroxidase 4, GPX4)和铁蛋白重链1 (ferritin heavy polypeptide 1, FTH1)基因表达水平。上述结果表明,LPS诱导奶牛乳腺上皮细胞的炎症反应和脂质过氧化,樱黄素通过激活Nrf2和抑制NF-κB来抑制脂质过氧化,进而减少LPS诱导的BMECs的炎性反应,从而达到细胞保护作用。本研究为樱黄素减轻LPS诱导的BMECs炎性反应和氧化损伤提供了理论支持,对于保护奶牛乳腺健康提供了新思路。

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	姚端阳
	兰政慧
	王若冰
	丰艳妮
	付开强
	李华涛
	刘锡武
	曹雷

关键词 ：奶牛乳腺上皮细胞(BMECs), 樱黄素, 脂质过氧化, 脂多糖(LPS), 核转录相关因子(Nrf2), 核因子κ, B (NF-κ, B)

Abstract：Mastitis has a significant impact on the health and milk production of dairy cows (Bos taurus), resulting in reduced quality of dairy products and significant economic losses to the entire dairy industry. Prunetin (Pru) had biological activities such as anti allergic and anti-inflammatory properties. The aim of this study was to investigate the effects and mechanisms of prunetin on the inflammatory response and lipid peroxidation induced by lipopolysaccharide in bovine mammary epithelial cells (BMECs). The experiment was divided into control (CK) group, lipopolysaccharide (LPS) group, and lipopolysaccharide plus prunetin (LPS+Pru) groups at concentrations of 10, 20, 40, and 80 μmol/L, as well as a lipopolysaccharide plus ferrostatin-1 (Fer-1)(LPS+Fer-1) group. The expression levels of inflammatory cytokines, nuclear factor κB (NF-κB), and nuclear factor erythroid 2-related factor 2 / Kelch-like ECH-associated protein 1 (Nrf2/Keap1) proteins were detected in each group. The results showed that LPS significantly upregulated the gene and protein levels of inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1β), IL-6, and IL-8. LPS also increased the content of Fe²⁺, reactive oxygen species (ROS), and malondialdehyde (MDA). Prunetin downregulated the genes of TNF-α, IL-1β, IL-6, IL-8, and their protein expression, upregulated the gene of Nrf2 and its protein expression, inhibited the NF-κB signaling pathway, and reduced cell death. Additionally, prunetin decreased the levels of Fe²⁺, ROS, and MDA while increasing the gene expression levels of glutathione peroxidase 4 (GPX4) and ferritin heavy polypeptide 1 (FTH1).These findings suggested that LPS induced inflammatory responses and lipid peroxidation in BMECs. Prunetin inhibited lipid peroxidation by activating Nrf2 and inhibiting NF-κB,and reducing the inflammatory response of BMECs induced by LPS, thereby exerting cytoprotective effects. This study provides theoretical support for the reduction of LPS induced inflammatory response and oxidative damage in BMECs by prunetin, and provides new ideas for protecting breast health in B. taurus.

Key words： Bovine mammary epithelial cells (BMECs) Prunetin Lipid peroxidation Lipopolysaccharide (LPS) NF-E2-related factor 2 (Nrf2) Nuclear factor κB (NF-κB)

收稿日期: 2024-01-08

中图分类号： S857.2

基金资助:山东省高等学校青创科技支持计划(2021KJ102); 山东省自然科学基金(ZR2023MC027; ZR2021MC111); 中央引导地方科技发展专项科技特派员行动计划(23-1-3-7-zyyd-nsh)

通讯作者: * huataoli@foxmail.com;caolei8004@163.com

引用本文:

姚端阳, 兰政慧, 王若冰, 丰艳妮, 付开强, 李华涛, 刘锡武, 曹雷. 樱黄素通过Nrf2通路抑制脂多糖诱导的奶牛乳腺上皮细胞炎症反应和脂质过氧化[J]. 农业生物技术学报, 2024, 32(7): 1554-1563.
YAO Duan-Yang, LAN Zheng-Hui, WANG Ruo-Bing, FENG Yan-Ni, FU Kai-Qiang, LI Hua-Tao, Liu Xi-Wu, CAO Lei. Prunetin Inhibits LPS-induced Inflammatory Response and Lipid Peroxidation in Bovine Mammary Epithelial Cells Through the Nrf2 Pathway. 农业生物技术学报, 2024, 32(7): 1554-1563.

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https://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2024.07.008 或 https://journal05.magtech.org.cn/Jwk_ny/CN/Y2024/V32/I7/1554

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