Tea polyphenols protect bovine mammary epithelial cells from hydrogen peroxide-induced oxidative damage in vitro
Tea polyphenols protect bovine mammary epithelial cells from hydrogen peroxide-induced oxidative damage in vitro
复制标题
茶多酚在体外保护牛乳腺上皮细胞免受过氧化氢诱导的氧化损伤
DOI：
10.1093/jas/sky278
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发表时间：
2018-10-01
影响因子：
3.3
通讯作者：
Loor, Juan J.
中科院分区：
文献类型：
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作者：
Ma, Yanfen;Zhao, Lei;Loor, Juan J.
围产期奶牛由于高代谢率和向泌乳过渡的生理适应特征而受到细胞内还原-氧化（氧化还原）平衡的改变。这些条件可以改变氧化应激状态。本研究旨在探讨茶多酚（TP）对过氧化氢（H2 O2）诱导的牛乳腺上皮细胞（BMEC）氧化损伤的保护作用。为了建立氧化应激，将分离的BMEC暴露于增加浓度的H2 O2（0、100、200、400、600、800和1，000 μ M）0、2、4、6、8、12和24 h。通过预处理BMEC 0、2、4、6、8、12和24 h，然后用H2 O2（600 μ M/培养孔）攻击6 h，评价TP剂量（0、60、80和100 μ g/mL）。将牛乳腺上皮细胞与或不与2，4-二硝基氯苯（DNCB）预孵育30分钟，然后与或不与TP（100 μ g/mL）再培养12小时，随后暴露于H2 O2（600 μ M/培养孔）。每个实验中每个处理有5个重复培养物。用600 μ M H2 O2/孔处理BMEC 6 h，可诱导BMEC氧化损伤，表明该体系可用于建立氧化应激模型。H2 O2（600 μ M每培养孔）曝光后，TP的浓度为100 μ g/mL，在12小时的孵育增加细胞活力，减少细胞内活性氧的积累，并增加核因子红细胞2相关因子2（NFE 2L 2）的丰度。此外，TP上调BMEC的NFE 2L 2和有丝分裂原活化蛋白激酶（MAPK）途径中基因的mRNA丰度。DNCB试验进一步证实，TP诱导NFE 2L 2和HMOX-1是通过持续上调MAPK 3/1 [以前称为细胞外调节激酶1/2]和MAPK 11/12/13/14（以前称为p38）的丰度介导的。总的来说，结果表明，TP对BMEC氧化还原平衡有有益的影响;它可以减少细胞氧化应激相关的损伤，并可能作为抗氧化剂对奶牛的氧化应激。
Periparturient dairy cows are subjected to altered intracellular reduction-oxidation (redox) balance due to the high metabolic rates and physiological adaptations characteristic of the transition into lactation. Such conditions could alter oxidative stress status. The objective of this study was to investigate the cytoprotective effects of tea polyphenols (TP) in cultured bovine mammary epithelial cells (BMEC) exposed to hydrogen peroxide (H2O2)-induced oxidative stress. To establish oxidative stress, isolated BMEC were exposed to increasing concentrations of H2O2 (0, 100, 200, 400, 600, 800, and 1,000 mu M) for 0, 2, 4, 6, 8, 12, and 24 h. Doses of TP (0, 60, 80, and 100 mu g/mL) were evaluated by pretreatment of BMEC for 0, 2, 4, 6, 8, 12 and 24 h, followed by an H2O2 (600 mu M per culture well) challenge for 6 h. Bovine mammary epithelial cells were preincubated for 30 min with or without 2,4-dinitrochloro-benzene (DNCB), then cultured with or without TP (100 mu g/mL) for another 12 h followed by H2O2 (600 mu M per culture well) exposure. There were 5 replicate cultures for each treatment in each experiment. Treatment with 600 mu M H2O2 per culture well for 6 h induced oxidative damage of BMEC, indicating this system could be used to establish an oxidative stress model. After H2O2 (600 mu M per culture well) exposure, a concentration of TP of 100 mu g/mL during a 12-h incubation increased cell viability, decreased intracellular reactive oxygen species accumulation, and increased the abundance of nuclear factor-erythroid 2-related factor 2 (NFE2L2). Furthermore, TP upregulated mRNA abundance of genes in the NFE2L2 and mitogen-activated protein kinase (MAPK) pathways of BMEC. The DNCB assay allowed further confirmation that the induction of NFE2L2 and HMOX-1 in response to TP was mediated through the sustained upregulation of the abundance of MAPK3/1 [formerly known as extracellular regulated kinases 1/2] and MAPK11/12/13/14 (formerly known as p38). Overall, results indicate that TP has beneficial effects on BMEC redox balance; it can reduce cellular oxidative stress-related injury and may potentially serve as an antioxidant against oxidative stress in dairy cows.