文献类型： 外文期刊

作者： Ma, Y. F. ¹ ; Wu, Z. H. ¹ ; Gao, M. ¹ ; Loor, J. J. ³ ;

作者机构： 1.Inner Mongolia Acad Agr & Anim Husb Sci, Inst Anim Nutr & Feed, Hohhot 010031, Peoples R China

2.Inner Mongolia Agr Univ, Coll Anim Sci, Hohhot 010018, Peoples R China

3.Univ Illinois, Dept Anim Sci, Div Nutr Sci, 328 Mumford Hall, Urbana, IL 61801 USA

关键词： nuclear factor (erythroid-derived 2)-like factor 2-antioxidant response element pathways; ataxia telangiectasia-mutated serine/threonine kinase; oxidative stress; bovine mammary epithelial cell

期刊名称：JOURNAL OF DAIRY SCIENCE （ 影响因子：4.034； 五年影响因子：4.354 ）

ISSN： 0022-0302

年卷期： 2018 年 101 卷 6 期

页码：

收录情况： SCI

摘要： Nuclear factor (erythroid-derived 2)-like factor 2 (NFE2L2, formerly Nrf2) is a transcription factor that binds to the antioxidant response element (ARE) in the upstream promoter region of various antioxidant-responsive genes. Hence, at least in nonruminants, the NFE2L2-ARE signaling pathway plays an important role in the cellular antioxidant defense system. Whether oxidative stress in bovine mammary epithelial cells alters NFE2L2 or the NFE2L2-ARE pathway is unclear. Therefore, the objective of this study was to examine the response in NFE2L2- and NFE2L2-ARE-related components in bovine mammary epithelial cells (BMEC) under oxidative stress. An in silico analysis to identify potential phosphorylation sites on NFE2L2 and the protein kinases was performed with Netphos 3.1 (http://www.cbs.dtu.dk/services/NetPhos/) and Scansite (http://scansite.rnit.edu ) software. Isolated BMEC were exposed to H2O2 (600 p,M) for 6 h to induce oxidative stress. In silico analysis revealed ataxia telangiectasia-mutated (ATM) serine/threonine kinase as a key kinase responsible for the phosphorylation of NFE2L2. Thus, after the 6 h incubation with H2O2, BMEC were transiently transfected with ATM-small interfering RNA (siRNA) 1, 2, or 3. Compared with the control, transfection with ATM-siRNA3 resulted in proliferation rates that were 60.7 and 36.2% lower with or without H2O2. In addition, production of reactive oxygen species and malondialdehyde increased markedly, but activities of superoxide dismutase, glutathione peroxidase, catalase, and glutathione-S-transferase decreased markedly in transfected cells without or with H2O2 compared with the control. Transfected cells had markedly lower protein and mRNA expression of NFE2L2 without or with H2O2 compared with the control. In addition, fluorescent activity of the ARE in transfected BMEC indicated that NFE2L2-driven transcriptional activation decreased under oxidative stress. Overall, results indicate that ATM is a physiologically relevant NFE2L2 kinase. Furthermore, inhibition of ATM activity can cause marked alterations in oxidative stress leading to cell death as a result of diminished capacity of BMEC to cope with H2O2-induced cytotoxicity. The relevance of this kinase in vivo merits further study.