Inhibiting Nuclear Factor Erythroid 2 Related Factor 2-mediated Autophagy in Bovine Mammary Epithelial Cells Induces Oxidative Stress in Response to Exogenous Fatty Acids

Overview

Journal J Anim Sci Biotechnol

Publisher Biomed Central

Date 2022 Apr 10

PMID 35397612

Authors

Renxu Chang

Xudong Sun

Hongdou Jia

Qiushi Xu

Zhihao Dong

Yan Tang

Shengbin Luo

Qianming Jiang

Juan J Loor

Chuang Xu

Affiliations

Soon will be listed here.

Abstract

Background: In early lactation, bovine mammary epithelial cells undergo serious metabolic challenges and oxidative stress both of which could be alleviated by activation of autophagy. Nuclear factor erythroid 2 related factor 2 (NFE2L2), a master regulator of cellular redox homeostasis, plays an important role in the regulation of autophagy and oxidative stress. Thus, the objective of this study was to investigate the role of NFE2L2-mediated autophagy on oxidative stress of bovine mammary epithelial cells in response to exogenous free fatty acids (FFA).

Results: Exogenous FFA induced linear and quadratic decreases in activities of glutathione peroxidase (GSH-Px), catalase (CAT), and superoxide dismutase (SOD), and increases in the contents of reactive oxygen species (ROS) and malondialdehyde (MDA). Protein abundance of LC3-phosphatidylethanolamine conjugate (LC3-II) and the number of autophagosomes and autolysosomes decreased in a dose-dependent manner, while protein abundance of p62 increased in cells challenged with FFA. Activation of autophagy via pre-treatment with Rap attenuated the FFA-induced ROS accumulation. Importantly, FFA inhibited protein abundance of NFE2L2 and the translocation of NFE2L2 into the nucleus. Knockdown of NFE2L2 by siRNA decreased protein abundance of LC3-II, while it increased protein abundance of p62. Furthermore, sulforaphane (SFN) pre-treatment attenuated the FFA-induced oxidative stress by activating NFE2L2-mediated autophagy.

Conclusions: The data suggested that NFE2L2-mediated autophagy is an important antioxidant mechanism in bovine mammary epithelial cells experiencing increased FFA loads.

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References

Liu H, Smith A, Ball S, Bao Y, Bowater R, Wang N . Sulforaphane promotes ER stress, autophagy, and cell death: implications for cataract surgery. J Mol Med (Berl). 2017; 95(5):553-564. PMC: 5403866. DOI: 10.1007/s00109-016-1502-4. View

Pajares M, Jimenez-Moreno N, Garcia-Yague A, Escoll M, de Ceballos M, Van Leuven F . Transcription factor NFE2L2/NRF2 is a regulator of macroautophagy genes. Autophagy. 2016; 12(10):1902-1916. PMC: 5079676. DOI: 10.1080/15548627.2016.1208889. View

Tanida I, Ueno T, Kominami E . LC3 and Autophagy. Methods Mol Biol. 2008; 445:77-88. DOI: 10.1007/978-1-59745-157-4_4. View

Feng T, Yin Q, Weng Z, Zhang J, Wang K, Yuan S . Rapamycin ameliorates neuropathic pain by activating autophagy and inhibiting interleukin-1β in the rat spinal cord. J Huazhong Univ Sci Technolog Med Sci. 2014; 34(6):830-837. DOI: 10.1007/s11596-014-1361-6. View

Song Y, Loor J, Li C, Liang Y, Li N, Shu X . Enhanced mitochondrial dysfunction and oxidative stress in the mammary gland of cows with clinical ketosis. J Dairy Sci. 2021; 104(6):6909-6918. DOI: 10.3168/jds.2020-19964. View

Tonelli C, Chio I, Tuveson D . Transcriptional Regulation by Nrf2. Antioxid Redox Signal. 2017; 29(17):1727-1745. PMC: 6208165. DOI: 10.1089/ars.2017.7342. View

Ornatowski W, Lu Q, Yegambaram M, Garcia A, Zemskov E, Maltepe E . Complex interplay between autophagy and oxidative stress in the development of pulmonary disease. Redox Biol. 2020; 36:101679. PMC: 7451718. DOI: 10.1016/j.redox.2020.101679. View

Sun X, Jia H, Xu Q, Zhao C, Xu C . Lycopene alleviates HO-induced oxidative stress, inflammation and apoptosis in bovine mammary epithelial cells via the NFE2L2 signaling pathway. Food Funct. 2019; 10(10):6276-6285. DOI: 10.1039/c9fo01922g. View

Glick D, Barth S, Macleod K . Autophagy: cellular and molecular mechanisms. J Pathol. 2010; 221(1):3-12. PMC: 2990190. DOI: 10.1002/path.2697. View

10.

Wu Y, Wang X, Guo H, Zhang B, Zhang X, Shi Z . Synthesis and screening of 3-MA derivatives for autophagy inhibitors. Autophagy. 2013; 9(4):595-603. PMC: 3627673. DOI: 10.4161/auto.23641. View

11.

Kim J, Sim H, Jung D, Lim E, Kim Y, Kim B . Wolf Extract Ameliorates Hepatic Steatosis through Regulation of Lipid Metabolism, Inhibition of ER Stress, and Activation of Autophagy via AMPK Activation. Int J Mol Sci. 2019; 20(19). PMC: 6801774. DOI: 10.3390/ijms20194801. View

12.

Sun X, Chang R, Tang Y, Luo S, Jiang C, Jia H . Transcription factor EB (TFEB)-mediated autophagy protects bovine mammary epithelial cells against HO-induced oxidative damage in vitro. J Anim Sci Biotechnol. 2021; 12(1):35. PMC: 7941962. DOI: 10.1186/s40104-021-00561-7. View

13.

Ma Y, Wu Z, Gao M, Loor J . Nuclear factor erythroid 2-related factor 2 antioxidant response element pathways protect bovine mammary epithelial cells against HO-induced oxidative damage in vitro. J Dairy Sci. 2018; 101(6):5329-5344. DOI: 10.3168/jds.2017-14128. View

14.

Sun X, Tang Y, Jiang C, Luo S, Jia H, Xu Q . Oxidative stress, NF-κB signaling, NLRP3 inflammasome, and caspase apoptotic pathways are activated in mammary gland of ketotic Holstein cows. J Dairy Sci. 2020; 104(1):849-861. DOI: 10.3168/jds.2020-18788. View

15.

Wang Q, Liang B, Shirwany N, Zou M . 2-Deoxy-D-glucose treatment of endothelial cells induces autophagy by reactive oxygen species-mediated activation of the AMP-activated protein kinase. PLoS One. 2011; 6(2):e17234. PMC: 3046135. DOI: 10.1371/journal.pone.0017234. View

16.

Gross J, Schwarz F, Eder K, van Dorland H, Bruckmaier R . Liver fat content and lipid metabolism in dairy cows during early lactation and during a mid-lactation feed restriction. J Dairy Sci. 2013; 96(8):5008-17. DOI: 10.3168/jds.2012-6245. View

17.

Tan W, Liao W, Peh H, Vila M, Dong J, Shen H . Andrographolide simultaneously augments Nrf2 antioxidant defense and facilitates autophagic flux blockade in cigarette smoke-exposed human bronchial epithelial cells. Toxicol Appl Pharmacol. 2018; 360:120-130. DOI: 10.1016/j.taap.2018.10.005. View

18.

Mohan N, Banik N, Ray S . Combination of N-(4-hydroxyphenyl) retinamide and apigenin suppressed starvation-induced autophagy and promoted apoptosis in malignant neuroblastoma cells. Neurosci Lett. 2011; 502(1):24-9. PMC: 3159706. DOI: 10.1016/j.neulet.2011.07.016. View

19.

Russell R, Yuan H, Guan K . Autophagy regulation by nutrient signaling. Cell Res. 2013; 24(1):42-57. PMC: 3879708. DOI: 10.1038/cr.2013.166. View

20.

Johansson I, Monsen V, Pettersen K, Mildenberger J, Misund K, Kaarniranta K . The marine n-3 PUFA DHA evokes cytoprotection against oxidative stress and protein misfolding by inducing autophagy and NFE2L2 in human retinal pigment epithelial cells. Autophagy. 2015; 11(9):1636-51. PMC: 4590664. DOI: 10.1080/15548627.2015.1061170. View