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

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

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G RX1、TRX1基因表达的调节作用。结果表明:1)G RX1、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40μg/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。

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, il- eum, 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 H202, 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 signifi- cantly 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 ex- pressions in those oxidative stress cells, and the optimal concentration of vitamin E was 40 ixg/mL. In conclu- sions, the expressions of GRX1 and TRX1 genes of Gaoligongshan pigs have obvious tissue- and gender-specif- ic manner; inducing those gene expressions might be an important way to relieve oxidative stress; the expres- sions of GRX1 and TRX1 genes and antioxidant ability can be improved by vitamin E supplementation at a suit- able concentration. [ Chinese Journal of Animal Nutrition, 2013, 25 （ 8 ） ： 1825-1835