title: Co- and Post-Treatment with Lysine Protects Primary Fish Enterocytes against Cu-Induced Oxidative Damage.
abstract: The aim of the work was primarily to explore the protective activity pathways of lysine against oxidative damage in fish in vivo and in enterocytes in vitro. First, grass carp were fed diets containing six graded levels of lysine (7.1-19.6 g kg-1 diet) for 56 days. Second, the enterocytes were treated with different concentrations of lysine (0-300 mg/L in media) prior to (pre-treatment), along with (co-treatment) or following (post-treatment) with 6 mg/L of Cu for 24 h. The results indicated that lysine improved grass carp growth performance. Meanwhile, lysine ameliorated lipid and protein oxidation by elevating the gene expression and activity of antioxidant enzymes (superoxide dismutase (SOD), glutathioneperoxidase (GPx), glutathione-S-transferase (GST) and reductase (GR)), and nuclear factor erythroid 2-related factor 2 (Nrf2) mRNA levels in fish intestine. The in vitro studies showed that co- and post-treatment with lysine conferred significant protection against Cu-induced oxidative damage in fish primary enterocytes as measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) OD values, along with alkaline phosphatase (ALP) and lactate dehydrogenase activities, and the depletion of protein carbonyl (PC), malondialdehyde (MDA) and 8-hydroxydeoxyguanosine contents. Moreover, lysine co-treatment decreased the activities and mRNA level of cellular SOD, GPx, GST and GR compared with the Cu-only exposed group. Gene expression of the signalling molecule Nrf2 showed the same pattern as that of SOD activity, whereas Kelch-like ECH-associated protein 1b (Keap1b) followed the opposite trend, indicating that co-treatment with lysine induced antioxidant enzymes that protected against oxidative stress through Nrf2 pathway. In addition, post-treatment with lysine increased proteasomal activity and blocked the Cu-stimulated increase in mRNA levels of GST and associated catalase (CAT) and GST activities (P<0.01 and P<0.001). GR activity and gene expression, and glutathione (GSH) content followed an opposite trend to GST activity (P<0.05). Thus, post-treatment of lysine elevated protein and DNA repair abilities and ameliorated the cellular redox state of enterocytes. The overall results suggest that lysine plays a significant role in the protection of fish intestine in vivo and in vitro through the induction of key antioxidant protection.
authors.affiliation: Animal Nutrition Institute, Sichuan Agricultural University, Chengdu, Sichuan, China.|Fish Nutrition and Safety Production University Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, China.
authors.forename: Xue-Yin
authors.initials: XY
authors.lastname: Li
authors.affiliation: Animal Nutrition Institute, Sichuan Agricultural University, Chengdu, Sichuan, China.|Fish Nutrition and Safety Production University Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, China.|Key Laboratory for Animal Disease-Resistance Nutrition of China Ministry of Education, Sichuan Agricultural University, Chengdu, Sichuan, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Animal Nutrition Institute, Sichuan Agricultural University, Chengdu, Sichuan, China.|Fish Nutrition and Safety Production University Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, China.|Key Laboratory for Animal Disease-Resistance Nutrition of China Ministry of Education, Sichuan Agricultural University, Chengdu, Sichuan, China.
authors.forename: Wei-Dan
authors.initials: WD
authors.lastname: Jiang
authors.affiliation: Animal Nutrition Institute, Sichuan Agricultural University, Chengdu, Sichuan, China.|Fish Nutrition and Safety Production University Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, China.|Key Laboratory for Animal Disease-Resistance Nutrition of China Ministry of Education, Sichuan Agricultural University, Chengdu, Sichuan, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Jiang
authors.affiliation: Animal Nutrition Institute, Sichuan Agricultural University, Chengdu, Sichuan, China.|Fish Nutrition and Safety Production University Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, China.|Key Laboratory for Animal Disease-Resistance Nutrition of China Ministry of Education, Sichuan Agricultural University, Chengdu, Sichuan, China.
authors.forename: Pei
authors.initials: P
authors.lastname: Wu
authors.affiliation: Animal Nutrition Institute, Sichuan Agricultural University, Chengdu, Sichuan, China.|Fish Nutrition and Safety Production University Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, China.|Key Laboratory for Animal Disease-Resistance Nutrition of China Ministry of Education, Sichuan Agricultural University, Chengdu, Sichuan, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Animal Nutrition Institute, Sichuan Academy of Animal Science, Chengdu, 610066, Sichuan, China.
authors.forename: Sheng-Yao
authors.initials: SY
authors.lastname: Kuang
authors.affiliation: Animal Nutrition Institute, Sichuan Academy of Animal Science, Chengdu, 610066, Sichuan, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Tang
authors.affiliation: Animal Nutrition Institute, Sichuan Academy of Animal Science, Chengdu, 610066, Sichuan, China.
authors.forename: Wu-Neng
authors.initials: WN
authors.lastname: Tang
authors.affiliation: Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, 430072, China.
authors.forename: Yong-An
authors.initials: YA
authors.lastname: Zhang
authors.affiliation: Animal Nutrition Institute, Sichuan Agricultural University, Chengdu, Sichuan, China.|Fish Nutrition and Safety Production University Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, China.|Key Laboratory for Animal Disease-Resistance Nutrition of China Ministry of Education, Sichuan Agricultural University, Chengdu, Sichuan, China.
authors.forename: Xiao-Qiu
authors.initials: XQ
authors.lastname: Zhou
authors.affiliation: Animal Nutrition Institute, Sichuan Agricultural University, Chengdu, Sichuan, China.|Fish Nutrition and Safety Production University Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, China.|Key Laboratory for Animal Disease-Resistance Nutrition of China Ministry of Education, Sichuan Agricultural University, Chengdu, Sichuan, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Feng
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D002347:Carps / Q000254:growth & development / Q000378:metabolism*; D003300:Copper / Q000633:toxicity*; D004260:DNA Repair / Q000187:drug effects; D020895:Enterocytes / Q000378:metabolism*; D008239:Lysine / Q000494:pharmacology*; D008315:Malondialdehyde / Q000378:metabolism; D051267:NF-E2-Related Factor 2 / Q000235:genetics / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D050050:Protein Carbonylation / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D012044:Regression Analysis; D015398:Signal Transduction / Q000187:drug effects; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism
chemical_list: D000975:Antioxidants; D051267:NF-E2-Related Factor 2; D012333:RNA, Messenger; D008315:Malondialdehyde; D003300:Copper; D010088:Oxidoreductases; D058570:TOR Serine-Threonine Kinases; D008239:Lysine
references.citation: Life Sci. 2003 Oct 24;73(23):3027-41
references.pmid: 14519451
references.citation: Clin Biochem. 2007 Nov;40(16-17):1252-8
references.pmid: 17915203
references.citation: Biochem J. 2011 Jan 1;433(1):51-63
references.pmid: 20961289
references.citation: PLoS One. 2014 Sep 19;9(9):e107670
references.pmid: 25238394
references.citation: Mech Ageing Dev. 2007 Mar;128(3):267-75
references.pmid: 17224177
references.citation: Fish Shellfish Immunol. 2014 Jun;38(2):374-82
references.pmid: 24751923
references.citation: Sci Total Environ. 2006 Dec 15;372(1):157-63
references.pmid: 17078999
references.citation: Science. 2009 Aug 14;325(5942):834-40
references.pmid: 19608861
references.citation: Semin Cell Dev Biol. 2012 Sep;23(7):729-37
references.pmid: 22484611
references.citation: Eur J Biochem. 1994 Feb 15;220(1):193-200
references.pmid: 8119287
references.citation: PLoS One. 2014 Dec 16;9(12):e115129
references.pmid: 25514235
references.citation: J Biol Chem. 2005 Jan 28;280(4):2737-44
references.pmid: 15550386
references.citation: Acta Histochem. 2012 Dec;114(8):785-96
references.pmid: 22341561
references.citation: Food Chem Toxicol. 2008 Sep;46(9):3015-22
references.pmid: 18601970
references.citation: Mol Cell Biol. 2003 Nov;23(22):8137-51
references.pmid: 14585973
references.citation: Toxicology. 2005 Jul 1;211(1-2):26-35
references.pmid: 15863245
references.citation: Free Radic Biol Med. 2001 Jun 1;30(11):1191-212
references.pmid: 11368918
references.citation: PLoS One. 2014 May 01;9(5):e95486
references.pmid: 24788257
references.citation: Arch Environ Contam Toxicol. 2010 Apr;58(3):757-64
references.pmid: 19789906
references.citation: PLoS One. 2012;7(10):e45870
references.pmid: 23056222
references.citation: Mol Cell Biol. 2004 Dec;24(24):10941-53
references.pmid: 15572695
references.citation: Meat Sci. 2009 Sep;83(1):104-12
references.pmid: 20416786
references.citation: Gen Comp Endocrinol. 2007 May 15;152(1):39-46
references.pmid: 17400223
references.citation: Eur J Biochem. 2004 Nov;271(21):4298-306
references.pmid: 15511236
references.citation: J Biol Chem. 2004 Jul 23;279(30):31556-67
references.pmid: 15143058
references.citation: Anal Biochem. 1976 May 7;72:248-54
references.pmid: 942051
references.citation: Diabetes. 1997 Nov;46(11):1733-42
references.pmid: 9356019
references.citation: J Ethnopharmacol. 2010 Feb 3;127(2):424-32
references.pmid: 19857565
references.citation: J Cell Biochem. 2008 Apr 15;103(6):1925-37
references.pmid: 17955498
references.citation: Fish Physiol Biochem. 2014 Jun;40(3):659-71
references.pmid: 24174167
references.citation: PLoS One. 2010 Jan 13;5(1):e8673
references.pmid: 20084273
references.citation: EMBO J. 2006 Aug 9;25(15):3605-17
references.pmid: 16888629
references.citation: Aquat Toxicol. 2011 Oct;105(3-4):543-51
references.pmid: 21924699
references.citation: PLoS One. 2014 Nov 25;9(11):e113712
references.pmid: 25423485
references.citation: Toxicology. 2000 Nov 16;153(1-3):83-104
references.pmid: 11090949
references.citation: Free Radic Biol Med. 2008 Jan 15;44(2):153-9
references.pmid: 18191751
references.citation: Comp Biochem Physiol C Toxicol Pharmacol. 2004 Apr;137(4):335-42
references.pmid: 15228951
references.citation: PLoS One. 2012;7(5):e37709
references.pmid: 22629446
references.citation: PLoS One. 2013 Aug 01;8(8):e70215
references.pmid: 23936392
references.citation: Chem Biol. 2010 Jan 29;17(1):75-85
references.pmid: 20142043
references.citation: Mol Aspects Med. 2003 Aug-Oct;24(4-5):167-75
references.pmid: 12892994
references.citation: J Nutr. 1998 Mar;128(3):606-14
references.pmid: 9482771
references.citation: Amino Acids. 2003 Dec;25(3-4):249-57
references.pmid: 14661088
references.citation: IUBMB Life. 2000 Oct-Nov;50(4-5):279-89
references.pmid: 11327322
references.citation: PLoS One. 2013 Jul 26;8(7):e69974
references.pmid: 23922879
references.citation: PLoS One. 2014 Jul 02;9(7):e100685
references.pmid: 24988078
references.citation: J Fish Dis. 2009 Sep;32(9):755-66
references.pmid: 19490393
references.citation: J Am Coll Nutr. 2003 Feb;22(1):18-35
references.pmid: 12569111
references.citation: Eur J Biochem. 1993 Jul 15;215(2):213-9
references.pmid: 7688300
references.citation: Int J Cell Biol. 2010;2010:632739
references.pmid: 20976254
references.citation: Aquat Toxicol. 2013 May 15;132-133:100-10
references.pmid: 23474319
references.citation: Science. 1983 Oct 21;222(4621):330-2
references.pmid: 6623079
references.citation: Hum Reprod Update. 2001 Mar-Apr;7(2):175-89
references.pmid: 11284661
references.citation: Mutat Res. 2004 Sep;567(1):1-61
references.pmid: 15341901
references.citation: Toxicol Pathol. 2002 Nov-Dec;30(6):620-50
references.pmid: 12512863
references.citation: Methods. 2001 Dec;25(4):402-8
references.pmid: 11846609
references.citation: Fish Shellfish Immunol. 2013 Oct;35(4):1235-9
references.pmid: 23973846
references.citation: Curr Opin Pharmacol. 2007 Aug;7(4):381-91
references.pmid: 17662654
references.citation: EMBO J. 2009 Feb 18;28(4):359-71
references.pmid: 19153599
references.citation: J Agric Food Chem. 2007 Oct 17;55(21):8458-64
references.pmid: 17883254
references.citation: Free Radic Biol Med. 2008 Apr 15;44(8):1617-23
references.pmid: 18279678
references.citation: Methods Enzymol. 1984;105:121-6
references.pmid: 6727660
references.citation: Cell Mol Biol (Noisy-le-grand). 2000 May;46(3):657-62
references.pmid: 10872752
references.citation: Am J Physiol Regul Integr Comp Physiol. 2001 Jan;280(1):R100-7
references.pmid: 11124139
references.citation: PLoS One. 2014 Apr 22;9(4):e95855
references.pmid: 24755687
references.citation: J Pharm Pharmacol. 2008 Jun;60(6):761-70
references.pmid: 18498713