动物营养学报 >

2013 , Vol. 25 >Issue 8: 1825 - 1835

DOI: https://doi.org/10.3969/j.issn.1006-267x.2013.08.020

分子营养 Molecular Nutrition

谷氧还蛋白1和硫氧还蛋白1基因在高黎贡山猪不同组织中表达规律及维生素E对其在氧化应激细胞中表达的影响

  • 安清聪 ,
  • 张春勇 ,
  • 李美荃 ,
  • 陈克嶙 ,
  • 郭荣富
展开
  • 云南农业大学,云南省动物营养与饲料重点实验室,昆明 650201

收稿日期: 2013-02-01

  网络出版日期: 2013-07-15

基金资助

云南现代农业生猪产业技术体系——生猪营养(A3007245);云南省重大科技专项项目(2012ZA018)

收起

Glutaredoxin 1 and Thioredoxin 1: Gene Expression Characteristics in Different Tissues and Effects of Vitamin E on Gene Expressions in Oxidative Stress Cells of Yunnan Gaoligongshan Pigs

  • AN Qingcong ,
  • ZHANG Chunyong ,
  • LI Meiquan ,
  • CHEN Kelin ,
  • GUO Rongfu
Expand
  • Yunnan Key Laboratory of Animal Nutrition and Feeds, Yunnan Agricultural University, Kunming 650201, China

Received date: 2013-02-01

  Online published: 2013-07-15

Fold

摘要

本试验研究云南特有的高黎贡山猪不同组织中谷氧还蛋白1(GRX1)、硫氧还蛋白1(TRX1)基因的表达规律,并探讨维生素E对高黎贡山猪氧化应激细胞中该基因表达的调节作用.采用实时荧光定量PCR(RT-qPCR)方法检测高黎贡山猪脑垂体、下丘脑、甲状腺、胸腺、胰腺、生殖腺、十二指肠、空肠、回肠、皮肤、肝脏11种组织中GRX1、TRX1基因表达;以过氧化氢(H2O2)为氧化应激源,建立猪胎儿皮肤成纤维细胞(PFSF)氧化应激模型,研究维生素E对GRX1、TRX1基因表达的调节作用.结果表明:1)GRX1、TRX1基因在高黎贡山猪被检11种组织中均有表达,基因表达谱为肝脏中GRX1、TRX1基因表达量最高,其次为皮肤、空肠,而胰腺和内分泌腺各组织中的表达量不高.母猪胸腺、生殖腺、回肠、肝脏中GRX1基因表达量显著高于公猪(P<0.05),胰腺中GRX1基因表达量极显著高于公猪(P<0.01);母猪垂体、胸腺、胰腺、生殖腺和肝脏中TRX1基因表达量显著高于公猪(P<0.05).2)细胞培养结果表明,受到H2O2刺激时,氧化应激细胞中TRX1基因产生过表达(P<0.05),维生素E对GRX1、TRX1基因表达具有明显的调节作用,其适宜浓度为40 μg/mL.综上所述,高黎贡山猪GRX1、TRX1基因的表达存在明显的性别和组织特异性;诱导猪体内GRX1、TRX1基因的表达可能是缓解机体氧化应激的一种重要方式;添加适宜浓度的维生素E可以提高氧化应激细胞中GRX1、TRX1基因表达,增强机体的抗氧化能力.

关键词： GRX1; TRX1; 基因表达; 高黎贡山猪; 氧化应激; 维生素E

本文引用格式

安清聪 , 张春勇 , 李美荃 , 陈克嶙 , 郭荣富 . 谷氧还蛋白1和硫氧还蛋白1基因在高黎贡山猪不同组织中表达规律及维生素E对其在氧化应激细胞中表达的影响[J]. 动物营养学报, 2013 , 25(8) : 1825 -1835 . DOI: 10.3969/j.issn.1006-267x.2013.08.020

Abstract

This experiment was conducted to study glutaredoxin 1 (GRX1) and Thioredoxin 1 (TRX1) gene expression characteristics in different tissues and investigate the regulation of vitamin E on the expressions of those two genes in oxidative stress cells of Yunnan Gaoligongshan pigs. The expressions of GRX1 and TRX1 genes in 11 tissues (hypophysis, hypothalamus, thyroid, thymus, pancreas, germen, duodenum, jejunum, ileum, skin and liver) were determined by the method of real time-quantitative PCR (RT-qPCR); pig fetal skin fibroblasts (PFSF) oxidative stress model was established induced by H2O2, and the regulation of vitamin E on GRX1 and TRX1 gene expressions in oxidative stress cells was explored. The results showed as follows: 1) GRX1 and TRX1 gene expressions were detected in all tissues, the highest expressional levels of GRX1 and TRX1 were in liver, and then skin and jejunum, and the lowest expressional levels was in pancreas and other endocrine glands; the expression level of GRX1 gene in thymus, germen, ileum and liver of sows was significantly higher than that of boars (P<0.05), and that in pancreas of sows was extremely higher than that of boars (P<0.01). The expression level of TRX1 gene in hypophysis, thymus, pancreas, germen and liver of sows was significantly higher than that of boars (P<0.05). 2) The results of cell culture indicated that TRX1 gene was over expressed in oxidative stress cells (P<0.05). Vitamin E regulated GRX1 and TRX1 gene expressions in those oxidative stress cells, and the optimal concentration of vitamin E was 40 μg/mL. In conclusions, the expressions of GRX1 and TRX1 genes of Gaoligongshan pigs have obvious tissue- and gender-specific manner; inducing those gene expressions might be an important way to relieve oxidative stress; the expressions of GRX1 and TRX1 genes and antioxidant ability can be improved by vitamin E supplementation at a suitable concentration.

Key words： GRX1; TRX1; gene expression; Gaoligongshan pigs; oxidative stress; vitamin E

参考文献

[1] 云南省畜牧局.云南省家畜家禽品种志[M].昆明:云南科技出版社,1987.

[2] 赵桂英,严达伟,连林生,等.高黎贡山猪遗传资源现状的调查[J].云南农业大学学报,2010,25(2):219-225.

[3] 连林生,屈再久,李琦华,等.高黎贡山猪[J].养猪,2004(2):24-25.

[4] HOLMGREN A.Antioxidant function of thioredoxin and glutaredoxin systems[J].Antioxidants & Redox Signaling,2000,2(4):811-820.  

[5] KERR C A,BUNTER K L,SEYMOUR R,et al.The heritability of the expression of two stress-regulated gene fragments in pigs[J].Journal of Animal Science,2005,83:1753-1765.

[6] TIM E,MARIO V P,PEELMAN L J,et al.Development of a new set of reference genes for normalization of real-time RT-PCR data of porcine backfat and longissimus dorsi muscle,and evaluation with PPARGC1A[J].BMC Biotechnology,2006,6:41.

[7] PFAFFL M W.A new mathematical model for relative quantification in real-time RT-PCR[J].Nucleic Acids Research,2001,29(9):e45.

[8] GALLOGLY M M,SHELTON M D,QANUNGO S,et al.Glutaredoxin regulates apoptosis in cardiomyocytes via NFκB targets Bcl-2 and Bcl-xL:implications for cardiac aging[J].Antioxidants & Redox Signaling,2010,12(12):1339-1353.  

[9] SHERLOCK S,DOOLEY J.Diseases of the liver and biliary system[M].11th ed.Oxford:Blackwell Science Ltd.,2007.

[10] SATDARSHAN P S M.Molecular pathology of liver diseases[M].New York:Springer-Verlag New York Inc.,2010.

[11] MORETO M,PEREZ A B.Dietary plasma proteins,the intestinal immune system,and the barrier functions of the intestinal mucosa[J].Journal of Animal Science,2009,87:E92-E100.

[12] O’NEIL M R,LARDY G P,REYNOLDS L P,et al.Impacts of linseed meal and estradiol-17β on mass,cellularity,angiogenic factors,and vascularity of the jejunum[J].Journal of Animal Science,2008,86:3014-3022.

[13] ANDERSON R R,PARRISH J A.The optics of human skin[J].Journal of Investigative Dermatology,1981,77:13-19.

[14] NESTLE F O,MEGLIO P D,JIAN Z Q.et al.Skin immune sentinels in health and disease[J].Nature Reviews Immunology,2009,9(10):679-691.

[15] IVARSSON R,QUINTENS R,DEJONGHE S,et al.Regulatory role of NADPH,thioredoxin, and glutaredoxin[J].Diabetes,2005,54(7):2132-2142.  

[16] BAKER A F,DRAGOVICHA T,TATEA W R,et al.The antitumor thioredoxin-1 inhibitor PX-12 (1-methylpropyl 2-imidazolyl disulfide) decreases thioredoxin-1 and VEGF levels in cancer patient plasma[J].Journal of Laboratory and Clinical Medicine,2006,147(2):83-90.  

[17] NAKAMURA H,BAI J,NISHINAKA Y,et al.Expression of thioredoxin and glutaredoxin,redox-regulating proteins,in pancreatic cancer[J].Cancer Detection and Prevention,2000,24(1):53-60.

[18] RAVID D,MUNIYAPPA H,DAS K C.Endogenous thioredoxin is required for redox cycling of anthracyclines and p53-dependent apoptosis in cancer cells[J].The Journal of Biological Chemistry,2005,280(48):40084-40096.  

[19] 刘莹莹.松江鲈(Trachidermus fasciatus)抗氧化相关基因的克隆、表达与功能分析 .博士学位论文.济南:山东大学,2012.

[20] RHEE S G.H2O2,a necessary evil for cell signaling[J].Science,2006,312:1882-1883.

[21] 邹朝霞.GRX1过表达在H2O2诱导的p38MAPK途径中的作用 .硕士学位论文.哈尔滨:哈尔滨医科大学,2006.

[22] NORIHIKO K,YASUYUKI I,JUNJI Y, et al.Redox-sensing release of human thioredoxin from T lymphocytes with negative feedback loops[J].The Journal of Immunology,2004,172:442-448.

[23] SANYAL A J,CHALASANI N,KOWDLEY K V,et al.Pioglitazone,vitamin E,or placebo for nonalcoholic steatohepatitis[J].The New England Journal of Medicine,2010,362(18):1675-1685.  

[24] 李彦,杨在宾,杨维仁,等.日粮维生素A和维生素E水平对肉鸡抗氧化和免疫性能的影响[J].动物营养学报,2008,20(4):417-422.

[25] 蒋守群,周桂莲,蒋宗勇,等.饲粮维生素E水平对43-63日龄黄羽肉鸡肉品质和抗氧化功能的影响[J].动物营养学报,2012,24(4):646-653.

[26] NAVA M,QUIROZ Y,VAZIRI N,et al.Melatonin reduces renal interstitial inflammation and improves hypertension in spontaneously hypertensive rats[J].American Journal of Physiology Renal Physiology,2003,284:F447-F454.

[27] 丛义梅.维生素E抗紫外线诱导的北京油鸡成纤维细胞凋亡作用的研究 .硕士学位论文.北京:中国农业科学院,2008.
Options
文章导航

/

〈 〉

    联系我们

  • 地址：北京市海淀区圆明园西路2号
    　　　中国农业大学动科动医大楼153室
  • 邮编：100193
  • 电话：010－62817823
  • 网址：http://www.chinajan.com
  • E-mail：cjan@chinajan.com
京公网安备 11010802041926号    京ICP备17050989号-2