Dataset Information

Effects of High-Grain Diet With Buffering Agent on the Hepatic Metabolism in Lactating Goats.

ABSTRACT: To gain insight on the effects of a high-grain diet with buffering agent on liver metabolism and the changes of plasma biochemical parameters and amino acids in hepatic vein and portal vein, commercial kit and high performance liquid chromatography (HPLC) were applied to determine the concentration of amino acids of hepatic vein and portal vein blood samples, quantitative real-time PCR and comparative proteomic approach was employed to investigate proteins differentially expressed in liver in lactating dairy goats feeding high-grain diet with buffering agent or only high-grain diet. Results showed that feeding high-grain diet with buffering agent to lactating dairy goats could outstanding increase amino acid content of Gln (p < 0.01), and the amino acid contents of Arg and Tyr in BG were significantly higher (p < 0.05) than that in HG. After adding the buffering agent, the metabolism of amino acids in the liver were changed and most of the amino acids were increasingly synthesized and decreasingly consumed in the liver. In addition, 46 differentially expressed protein spots (?1.5-fold changed) were detected in buffering group vs. control group using 2-DE technique and MALDI-TOF/TOF proteomics analyzer. Of these, 24 proteins showed increased expression and 22 proteins showed decreased expression in the buffer group vs. control group. Data on Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis reveals that the high-grain diet with buffering agent alter the expression of proteins related to amino acids metabolism and glycometabolism. In addition, the results conclude that feeding high-grain diet with buffering agent can strengthen anti-oxidant capacity, stress ability, slow down urea metabolism, and alter amino acid metabolism as well as glycometabolism in the liver through different detection methods including proteomic analysis, real-time PCR analysis and biochemical analysis.

SUBMITTER: He M

PROVIDER: S-EPMC6548822 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Effects of High-Grain Diet With Buffering Agent on the Hepatic Metabolism in Lactating Goats.

He Meilin M Li Lin L Wang Huanhuan H Yan Shuping S Zhang Yuanshu Y

Frontiers in physiology 20190529

To gain insight on the effects of a high-grain diet with buffering agent on liver metabolism and the changes of plasma biochemical parameters and amino acids in hepatic vein and portal vein, commercial kit and high performance liquid chromatography (HPLC) were applied to determine the concentration of amino acids of hepatic vein and portal vein blood samples, quantitative real-time PCR and comparative proteomic approach was employed to investigate proteins differentially expressed in liver in la ...[more]

PMID: 31191354

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Project 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 5? (ATP5?), 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 ?-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.

Project description:BackgroundIn ruminants, lower ruminal pH causes massive disruption of ruminal epithelial structure during periods of feeding high-concentrate diets. However, the influence of excessive organic fatty acids in the lumen of hindgut on the epithelial structure is unclear. In this study, twelve mid-lactating 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 weeks. The colonic epithelial structure was detected by HE staining and transmission electron microscopy (TEM), and the apoptotic status of epithelial cells was estimated by TUNEL method and caspase activities.ResultsHC goats showed higher level of free lipopolysaccharide (LPS) in rumen fluid (p < 0.01) but not in colonic digesta (p > 0.05), and higher total volatile fatty acid (VFA) concentrations in rumen fluid (p < 0.05) and in colonic digesta (p < 0.01), and higher content of starch in colonic digesta (p < 0.05) compared to LC goats. HC goats demonstrated profound alterations in the colonic epithelial structure and tight junctions (TJ), apparently due to damage of the epithelium with widened TJs space and nuclear breakdown and mitochondrial swelling. HC goats showed higher level of apoptosis in the colonic epithelium with higher proportion of TUNEL-positive apoptotic cells and increases of caspase-3 and -3/7 activities, as well as the lower ratio of bcl-2/bax mRNA expression in the colonic mucosa (p < 0.05). However, β-defense mRNA was significantly down-regulated in the colonic mucosa of HC goats compared to LC (p < 0.05). HC goats showed higher level of TJ proteins including claudin-1 and claudin-4 in the colonic mucosa than LC (p < 0.05). Neither free LPS content in the colonic digesta nor NF-κ B protein expression in tissues showed significant difference between HC and LC goats (p > 0.05).ConclusionsOur results reveal that long-term feeding HC diet to lactating goats causes severe damages to the colonic mucosa barrier associated with activating cells apoptosis.

Project description:BackgroundIt is well known that feeding a high concentrate (HC) diet to lactating ruminants likely induces subacute ruminal acidosis (SARA) and leads to a decrease in milk fat production. However, the effects of feeding a HC diet for long periods on milk fatty acids composition and the mechanism behind the decline of milk fat still remains poorly understood. The aim of this study was to investigate the impact of feeding a HC diet to lactating dairy goats on milk fat yield and fatty acids composition with an emphasis on the mechanisms underlying the milk fat depression. Seventeen mid-lactating dairy goats were randomly allocated to three groups. The control treatment was fed a low-concentrate diet (35% concentrate, n = 5, LC) and there were two high-concentrate treatments (65% concentrate, HC), one fed a high concentrate diet for a long period (19 wks, n = 7, HL); one fed a high concentrate diet for a short period of time (4 wk, n = 5, HS). Milk fat production and fatty acids profiles were measured. In order to investigate the mechanisms underlying the changes in milk fat production and composition, the gene expression involved in lipid metabolism and DNA methylation in the mammary gland were also analyzed.ResultsMilk production was increased by feeding the HC diet in the HS and HL groups compared with the LC diet (P < 0.01), while the percentage of milk fat was lower in the HL (P < 0.05) but not in the HS group. The total amount of saturated fatty acids (SFA) in the milk was not changed by feeding the HC diet, whereas the levels of unsaturated fatty acids (UFA) and monounsaturated fatty acids (MUFA) were markedly decreased in the HL group compared with the LC group (P < 0.05). Among these fatty acids, the concentrations of C15:0 (P < 0.01), C17:0 (P < 0.01), C17:1 (P < 0.01), C18:1n-9c (P < 0.05), C18:3n-3r (P < 0.01) and C20:0 (P < 0.01) were markedly lower in the HL group, and the concentrations of C20:0 (P < 0.05) and C18:3n-3r (P < 0.01) were lower in the HS group compared with the LC group. However, the concentrations of C18:2n-6c (P < 0.05) and C20:4n-6 (P < 0.05) in the milk fat were higher in the HS group. Real-time PCR results showed that the mRNA expression of the genes involved in milk fat production in the mammary gland was generally decreased in the HL and HS groups compared with the LC group. Among these genes, ACSL1, ACSS1 & 2, ACACA, FAS, SCD, FADS2, and SREBP1 were down-regulated in the mammary gland of the HL group (P < 0.05), and the expressions of ACSS2, ACACA, and FADS2 mRNA were markedly decreased in the HS goats compared with the LC group (P < 0.05). In contrast to the gene expression, the level of DNA methylation in the promoter regions of the ACACA and SCD genes was increased in the HL group compared with the LC group (P < 0.05). The levels of ACSL1 protein expression and FAS enzyme activity were also decreased in the mammary gland of the HL compared with the LC group (P < 0.05).ConclusionsLong-term feeding of a HC diet to lactating goats induced milk fat depression and FAs profile shift with lower MUFAs but higher SFAs. A general down-regulation of the gene expression involved in the milk fat production and a higher DNA methylation in the mammary gland may contribute to the decrease in milk fat production in goats fed a HC diet for long time periods.

Project description:The objective of this study was to reveal the effect of rumen degradable starch (RDS) on bile acid metabolism and liver transcription in dairy goats using metabolomics and transcriptomics. Eighteen Guanzhong dairy goats of a similar weight and production level (body weight = 45.8 ± 1.54 kg, milk yield = 1.75 ± 0.08 kg, and second parity) were randomly assigned to 3 treatment groups where they were fed a low RDS (LRDS, RDS = 20.52% DM) diet, medium RDS (MRDS, RDS = 22.15% DM) diet, or high RDS (HRDS, RDS = 24.88% DM) diet, respectively. The goats were fed with the experimental diets for 5 weeks. On the last day of the experiment, all goats were anesthetized, and peripheral blood and liver tissue samples were collected. The peripheral blood samples were used in metabolomic analysis and white blood cell (WBC) count, whereas the liver tissue samples were used in transcriptomic analysis. Based on the metabolomics results, the relative abundances of primary bile acids in the peripheral blood were significantly reduced in the group that was fed the HRDS diet (P < 0.05). The WBC count was significantly increased in the HRDS group compared with that in the LRDS and MRDS groups (P < 0.01), indicating that there was inflammation in the HRDS group. Transcriptomic analysis showed that 4 genes related to bile acid secretion (genes: MDR1, RXRα, AE2, SULT2A1) were significantly downregulated in the HRDS group. In addition, genes related to the immune response were upregulated in the HRDS group, suggesting the HRDS diet induced a hepatic inflammatory response mediated by lipopolysaccharides (LPS) (gene: LBP), activated the Toll-like receptor 4 binding (genes: S100A8, S100A9) and the NF-kappa B signaling pathway (genes: LOC106503980, LOC108638497, CD40, LOC102180880, LOC102170970, LOC102175177, LBP, LOC102168903, LOC102185461, LY96 and CXCL8), triggered inflammation and complement responses (genes: C1QB, C1QC, and CFD). The HRDS diet induced a hepatic inflammatory response may be mediated by activating the Toll-like receptor 4 binding and NF-kappa B signaling pathway after free LPS entered the liver. The changes of bile acids profile in blood and the down-regulation of 4 key genes (MDR1, RXRα, AE2, SULT2A1) involved in bile secretion in liver are probably related to liver inflammation.

Dataset Information

Effects of High-Grain Diet With Buffering Agent on the Hepatic Metabolism in Lactating Goats.

Publications

Effects of High-Grain Diet With Buffering Agent on the Hepatic Metabolism in Lactating Goats.

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