维生素C对建鲤肠上皮细胞氧化损伤保护作用的研究

刘扬; 池磊; 冯琳; 姜俊; 周小秋

doi:10.3969/j.issn.1006-267x.2012.08.014

动物营养学报 >

2012 , Vol. 24 >Issue 8: 1503 - 1511

DOI: https://doi.org/10.3969/j.issn.1006-267x.2012.08.014

水产营养 Fish Nutrition

维生素C对建鲤肠上皮细胞氧化损伤保护作用的研究

刘扬 ,
池磊 ,
冯琳 ,
姜俊 ,
周小秋

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1. 四川农业大学动物营养研究所,雅安 625014;
2. 鱼类营养与安全生产四川省高校重点实验室,雅安 625014;
3. 动物抗病营养教育部重点实验室,雅安 625014;
4. 成都三旺农牧股份有限公司,新津 621430

收稿日期: 2012-05-14

网络出版日期: 2012-08-03

基金资助

四川省教育厅重点项目(2009ZA070)

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Protective Effect of Vitamin C on Oxidative Damage in Intestinal Epithelial Cells of Jian Carp (Cyprinus carpio var. Jian)

LIU Yang ,
CHI Lei ,
FENG Lin ,
JIANG Jun ,
ZHOU Xiaoqiu

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1. Animal Nutrition Institute, Sichuan Agricultural University, Ya’an 625014, China;
2. Fish Nutrition and Safety Production University Key Laboratory of Sichuan Province, Ya’an 625014, China;
3. Key Laboratory for Animal Disease-Resistance Nutriton of China Ministy of Education, Ya’an 625014, China;
4. Chengdu Sunwind Agricaltural and Livestack Farming Co. , Ltd. , Xinjin 621430, China

Received date: 2012-05-14

Online published: 2012-08-03

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摘要

建鲤肠上皮细胞(IEC)在24孔培养板中原代培养72 h并更换无血清DMEM培养液继续培养12 h后,随机分为7组,每组4个重复。除其中1组作为空白对照外,另外6组均加入100 μmol/L的过氧化氢(H₂O₂)以建立IEC氧化应激模型。相同条件下继续培养16 h后,将培养液分别更换为含0(空白对照组)、0、4、10、16、22和28 mg/L维生素C的无血清DMEM培养液,相同条件下继续培养72 h后取样测定指标,以考察不同维生素C浓度对H₂O₂诱导的建鲤IEC氧化损伤的保护作用。结果显示:1)100 μmol/L H₂O₂处理导致IEC存活数量减少,仅形成一些小细胞集落,并显著降低IEC中超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶、谷胱甘肽硫转移酶和谷胱甘肽还原酶活性以及谷胱甘肽和蛋白质含量(P<0.05),提高IEC中丙二醛及蛋白质羰基含量以及培养液中乳酸脱氢酶活性(P<0.05),使建鲤IEC产生了氧化应激;2)维生素C显著提高IEC的抗羟自由基能力和抗超氧阴离子自由基能力(P<0.05),缓解由H₂O₂导致的建鲤IEC氧化损伤。综上可见,维生素C可提高建鲤IEC抗氧化能力,有效保护IEC免受氧化损伤。

关键词： 建鲤; 维生素C; 肠上皮细胞; 抗氧化能力

本文引用格式

刘扬 , 池磊 , 冯琳 , 姜俊 , 周小秋 . 维生素C对建鲤肠上皮细胞氧化损伤保护作用的研究[J]. 动物营养学报, 2012 , 24(8) : 1503 -1511 . DOI: 10.3969/j.issn.1006-267x.2012.08.014

Abstract

This experiment was conducted to investigate the protective effect of different concentrations of vitamin C on hydrogen peroxide (H₂O₂)-induced oxidative damage in intestinal epithelial cells (IEC) of Jian carp. In 24-well culture plates, the Jian carp IEC were primarily cultured for 72 h, and then continued to culture in serum-free DMEM culture medium. After 12 h further culture, the IEC were randomly divided into 7 groups with 4 replicates per group. One of the 7 groups was set as blank control, and the other groups were all supplemented with 100 μmol/L H₂O₂ in culture medium in order to establish oxidative stress model. After 16 h further culture in the same condition, the culture medium was changed into the DMEM culture medium which contained 0 (blank control group), 0, 4, 10, 16, 22 and 28 mg/L vitamin C, respectively. The samples were collected to measure the indices after 72 h vitamin C culture. The results showed as follows: 1) the 100 μmol/L H₂O₂ treatment led to less survival number and small colony of IEC; significantly decreased the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), glutathione S-transferase (GST) and glutathion reductase (GR), and reduced the contents of glutathione (GSH) and protein (P<0.05); significantly increased the contents of malonaldehyde (MDA) and protein carbonyl (PCA), and improved the lactate dehydrogenase (LDH) activity in culture medium (P<0.05); caused oxidative stress of Jian carp IEC. 2) Vitamin C significantly increased the anti-hydroxyl free radical ability (AHA) and anti-superoxide anion free radical ability (ASA) (P<0.05), and recovered the oxidation damage in IEC of Jian carp induced by H₂O₂. In summary, vitamin C can improve the antioxidant ability of the Jian carp IEC, and effectively protect the IEC from oxidative damage.

Key words： Jian carp; vitamin C; IEC; antioxidant ability

参考文献

[1] MITRA G,MUKHOPADHYAY P K,AYYAPPN S.Modulation of digestive enzyme activities during ontogeny of Labeo rohita larvae fed ascorbic acid enriched zooplankton[J].Comparative Biochemistry Physiology,2008,149A:341-350.

[2] KOHEN W,ROJAS-GARCIA C R,FIN R N,et al.Stimulatory effect of ingested protein and/or free amino acids on the secretion of the gastro-endocrine hormone cholecystokininand on tryptic activity,in early feeding herring larvae,Clupea harengus[J].Marine Biology,2002,140:1241-1247.

[3] DABROWSKI K,GUDERLEY H.Intermediary metabolism[M]//HALVER J E,HARDY R.Fish nutrition.New York:Academic Press,2002:309-365.

[4] 郑婷.维生素E对建鲤肠上皮细胞生长发育及抗氧化能力的影响 .硕士学位论文.雅安:四川农业大学,2007.

[5] LIU Y,CHI L,FENG L,et al.Effects of graded levels of dietary vitamin C on the growth,digestive capacity and intestinal microflora of juvenile Jian carp (Cyprinus carpio var.Jian)[J].Aquaculture Research,2011,42:534-548.

[6] 姜俊.谷氨酰胺对建鲤肠上皮细胞增殖和分化的影响 .硕士学位论文.雅安:四川农业大学,2005.

[7] AW T Y.Cellular redox:a modulator of intestinal epithelial cell proliferation[J].News in Physiological Sciences,2003,18:201-204.

[8] KOHEN R,NYSKA A.Oxidation of biological systems:oxidative stress phenomena,antioxidants,redox reactions,and methods for their quantification[J].Toxicologic Pathology,2002,30(6):620-650.

[9] 冯琳.大豆凝集素对鲤鱼肠上皮细胞增殖和分化的影响 .硕士学位论文.雅安:四川农业大学,2006.

[10] BASIVIREDDY J,JACOB M,BALASUBRAMANIAN K A,et al.Oral glutamine attenuates indomethacin induced small intestinal damage[J].Clinical Science,2004,107:281-289.

[11] GLISZCZY N ＇ SKA-S ＇ WIGLO A.Antioxidant activity of water soluble vitamins in the TEAC (trolox equivalent antioxidant capacity) and the FRAP (ferric reducing antioxidant power) assays[J].Food Chemistry,2006,96:131-136.

[12] 王秋林,王浩毅,王树人,等.氧化应激状态的评价[J].中国病理生理杂志,2005,21:2069-2074.

[13] 陈瑾.谷氨酰胺对鲤鱼肠上皮细胞抗氧化能力的影响 .硕士学位论文.雅安:四川农业大学,2008.

[14] BALTACIO G ˇ LU E,AKALIN F A,ALVER A,et al.Protein carbonyl levels in serum and gingival in patients with chronic periodontitis[J].Archives of Oral Biology,2008,53:716-722.

[15] BAI J,CEDERBAUM A I.Mitochondrial catalase and oxidative injury[J].Biological Signals and Receptors,2001,10:189-199.

[16] WINSTON G W,DI-GIULIO R T.Prooxidant and antioxidant mechanisms in aquatic organisms[J].Aquatic Toxicology,1991,19:137-161.

[17] ELIA A C,ANASTASI V,DRR A J M.Hepatic antioxidant enzymes and total glutathione of Cyprinus carpio exposed to three disinfectants, chlorine dioxide, sodium hypochlorite and peracetic acid for superficial water potabilization[J].Chemosphere,2006,64:1633-1641.

[18] 杨雨晗,王玉明,郑人源,等.维生素C对神经元细胞生长的时间剂量效应[J].营养学报,2009(5):460-464.

[19] HUANG J, MAY J M.Ascorbic acid spares alpha-tocopherol and prevents lipid peroxidation in cultured H4IIE liver cells[J].Molecular and Cellular Biochemistry,2003,247:171-176.

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