Comparative proteomic analysis of the effects of high-concentrate diet on the hepatic metabolism and inflammatory response in lactating dairy goats  

作　　者：Yongqian Duanmu Rihua Cong Shiyu Tao Jing Tian Haibo Dong Yuanshu Zhang Yingdong Ni Ruqian Zhao 

机构地区：[1]Key Laboratory of Animal Physiology & Biochemistry, Ministry of Agriculture,Nanjing Agricultural University [2]College of Veterinary Medicine, Northwest A & F University

出　　处：《Journal of Animal Science and Biotechnology》2016年第2期217-227,共11页畜牧与生物技术杂志（英文版）

基　　金：supported by National Nature Science Foundation of China (Project No. 31272470, No. 31572433);National Basic Research Program of China (Project No. 2011CB100802);A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD)

摘　　要：Background： To understand the impact of feeding a high-concentrate diet to mid-lactating goats for a long time on liver metabolism and inflammatory response, two dimensional polyacrylamide gel electrophoresis（2-DE） and real-time PCR method were employed to detect proteins differentially expressed in liver and their m RNAs expression in goats fed high concentrate diet（HC） or low concentrate diet（LC）. Twelve lactating dairy goats were randomly assigned to either a HC diet group（65 % concentrate of dry matter; n = 6） or a LC diet group（35 % concentrate of dry matter; n = 6） for 10 wk.Results： Twenty differentially expressed spots（≥2.0-fold changes） in the hepatic tissues were excised and successfully identified using MALDI TOF/TOF. Of these, 8 proteins were up-regulated, while the rest 12 proteins were down-regulated in HC goats compared to LC. Differential expressed proteins including alpha enolase 1（ENO1）, glutamate dehydrogenase 1（GLUD1）, glutathione S-transferase A1（GSTA1）, ATP synthase subunit 5β（ATP5β）, superoxide dismutase [Cu-Zn]（SOD1）, cytochrom c oxidase subunit Via（COX6A1） and heat shock protein 60（HSP60） were further verified by real-time PCR and/or western blot at m RNA or protein expression level. Consistent with the 2-DE results, a significant decrease of β-actin protein expression and SOD enzyme activity was observed in liver of HC goats（P 〈 0.05）, while ENO1 protein expression was significantly up-regulated in HC compared to LC goats（P 〈 0.05）. However, western blot analysis did not show a significant difference of hepatic HSP60 protein between HC and LC group, which did not match the decrease of HSP60 content detected by 2-DE analysis. Real-time PCR showed that glutathione S-transferase P1（GSTP1） and SOD1 m RNA expression was significantly decreased in liver of HC goats, while cytochrom c oxidase（COX3） and ATPase 8（ATP8） m RNAs expression were markedly increased compared to LC（P 〈 0.05）. Gene Ontology（GO）Background： To understand the impact of feeding a high-concentrate diet to mid-lactating goats for a long time on liver metabolism and inflammatory response, two dimensional polyacrylamide gel electrophoresis（2-DE） and real-time PCR method were employed to detect proteins differentially expressed in liver and their m RNAs expression in goats fed high concentrate diet（HC） or low concentrate diet（LC）. Twelve lactating dairy goats were randomly assigned to either a HC diet group（65 % concentrate of dry matter; n = 6） or a LC diet group（35 % concentrate of dry matter; n = 6） for 10 wk.Results： Twenty differentially expressed spots（≥2.0-fold changes） in the hepatic tissues were excised and successfully identified using MALDI TOF/TOF. Of these, 8 proteins were up-regulated, while the rest 12 proteins were down-regulated in HC goats compared to LC. Differential expressed proteins including alpha enolase 1（ENO1）, glutamate dehydrogenase 1（GLUD1）, glutathione S-transferase A1（GSTA1）, ATP synthase subunit 5β（ATP5β）, superoxide dismutase [Cu-Zn]（SOD1）, cytochrom c oxidase subunit Via（COX6A1） and heat shock protein 60（HSP60） were further verified by real-time PCR and/or western blot at m RNA or protein expression level. Consistent with the 2-DE results, a significant decrease of β-actin protein expression and SOD enzyme activity was observed in liver of HC goats（P 〈 0.05）, while ENO1 protein expression was significantly up-regulated in HC compared to LC goats（P 〈 0.05）. However, western blot analysis did not show a significant difference of hepatic HSP60 protein between HC and LC group, which did not match the decrease of HSP60 content detected by 2-DE analysis. Real-time PCR showed that glutathione S-transferase P1（GSTP1） and SOD1 m RNA expression was significantly decreased in liver of HC goats, while cytochrom c oxidase（COX3） and ATPase 8（ATP8） m RNAs expression were markedly increased compared to LC（P 〈 0.05）. Gene Ontology（GO）

关 键 词：High concentrate diet Lactating goats Liver Mitochondria Proteomics 

分 类 号：S823.5[农业科学—畜牧学]