Cargando…

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

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 i...

Descripción completa

Detalles Bibliográficos
Autores principales: Duanmu, Yongqian, Cong, Rihua, Tao, Shiyu, Tian, Jing, Dong, Haibo, Zhang, Yuanshu, Ni, Yingdong, Zhao, Ruqian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4744397/
https://www.ncbi.nlm.nih.gov/pubmed/26855776
http://dx.doi.org/10.1186/s40104-016-0065-0
_version_ 1782414482697355264
author Duanmu, Yongqian
Cong, Rihua
Tao, Shiyu
Tian, Jing
Dong, Haibo
Zhang, Yuanshu
Ni, Yingdong
Zhao, Ruqian
author_facet Duanmu, Yongqian
Cong, Rihua
Tao, Shiyu
Tian, Jing
Dong, Haibo
Zhang, Yuanshu
Ni, Yingdong
Zhao, Ruqian
author_sort Duanmu, Yongqian
collection PubMed
description 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 mRNAs 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 mRNA 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 mRNA expression was significantly decreased in liver of HC goats, while cytochrom c oxidase (COX3) and ATPase 8 (ATP8) mRNAs expression were markedly increased compared to LC (P < 0.05). Gene Ontology (GO) analysis revealed that HC diet resulted in altered expression of proteins related to catalytic and mitochondrial metabolism in the liver, and may increase the stress response with up-regulating the expression of differentiation 14 (CD14) cluster and serum amyloid A (SAA) as well as C-reactive protein (CRP) in the liver. CONCLUSIONS: These results suggest that feeding high concentrate diet to lactating goats for 10 wk leads to the activation of the inflammatory response, and decreases the anti-oxidant capacity, and subsequently impairs the mitochondrial function in the liver. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s40104-016-0065-0) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-4744397
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-47443972016-02-07 Comparative proteomic analysis of the effects of high-concentrate diet on the hepatic metabolism and inflammatory response in lactating dairy goats Duanmu, Yongqian Cong, Rihua Tao, Shiyu Tian, Jing Dong, Haibo Zhang, Yuanshu Ni, Yingdong Zhao, Ruqian J Anim Sci Biotechnol Research 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 mRNAs 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 mRNA 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 mRNA expression was significantly decreased in liver of HC goats, while cytochrom c oxidase (COX3) and ATPase 8 (ATP8) mRNAs expression were markedly increased compared to LC (P < 0.05). Gene Ontology (GO) analysis revealed that HC diet resulted in altered expression of proteins related to catalytic and mitochondrial metabolism in the liver, and may increase the stress response with up-regulating the expression of differentiation 14 (CD14) cluster and serum amyloid A (SAA) as well as C-reactive protein (CRP) in the liver. CONCLUSIONS: These results suggest that feeding high concentrate diet to lactating goats for 10 wk leads to the activation of the inflammatory response, and decreases the anti-oxidant capacity, and subsequently impairs the mitochondrial function in the liver. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s40104-016-0065-0) contains supplementary material, which is available to authorized users. BioMed Central 2016-02-06 /pmc/articles/PMC4744397/ /pubmed/26855776 http://dx.doi.org/10.1186/s40104-016-0065-0 Text en © Duanmu et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Duanmu, Yongqian
Cong, Rihua
Tao, Shiyu
Tian, Jing
Dong, Haibo
Zhang, Yuanshu
Ni, Yingdong
Zhao, Ruqian
Comparative proteomic analysis of the effects of high-concentrate diet on the hepatic metabolism and inflammatory response in lactating dairy goats
title Comparative proteomic analysis of the effects of high-concentrate diet on the hepatic metabolism and inflammatory response in lactating dairy goats
title_full Comparative proteomic analysis of the effects of high-concentrate diet on the hepatic metabolism and inflammatory response in lactating dairy goats
title_fullStr Comparative proteomic analysis of the effects of high-concentrate diet on the hepatic metabolism and inflammatory response in lactating dairy goats
title_full_unstemmed Comparative proteomic analysis of the effects of high-concentrate diet on the hepatic metabolism and inflammatory response in lactating dairy goats
title_short Comparative proteomic analysis of the effects of high-concentrate diet on the hepatic metabolism and inflammatory response in lactating dairy goats
title_sort comparative proteomic analysis of the effects of high-concentrate diet on the hepatic metabolism and inflammatory response in lactating dairy goats
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4744397/
https://www.ncbi.nlm.nih.gov/pubmed/26855776
http://dx.doi.org/10.1186/s40104-016-0065-0
work_keys_str_mv AT duanmuyongqian comparativeproteomicanalysisoftheeffectsofhighconcentratedietonthehepaticmetabolismandinflammatoryresponseinlactatingdairygoats
AT congrihua comparativeproteomicanalysisoftheeffectsofhighconcentratedietonthehepaticmetabolismandinflammatoryresponseinlactatingdairygoats
AT taoshiyu comparativeproteomicanalysisoftheeffectsofhighconcentratedietonthehepaticmetabolismandinflammatoryresponseinlactatingdairygoats
AT tianjing comparativeproteomicanalysisoftheeffectsofhighconcentratedietonthehepaticmetabolismandinflammatoryresponseinlactatingdairygoats
AT donghaibo comparativeproteomicanalysisoftheeffectsofhighconcentratedietonthehepaticmetabolismandinflammatoryresponseinlactatingdairygoats
AT zhangyuanshu comparativeproteomicanalysisoftheeffectsofhighconcentratedietonthehepaticmetabolismandinflammatoryresponseinlactatingdairygoats
AT niyingdong comparativeproteomicanalysisoftheeffectsofhighconcentratedietonthehepaticmetabolismandinflammatoryresponseinlactatingdairygoats
AT zhaoruqian comparativeproteomicanalysisoftheeffectsofhighconcentratedietonthehepaticmetabolismandinflammatoryresponseinlactatingdairygoats