Journal
ASIAN-AUSTRALASIAN JOURNAL OF ANIMAL SCIENCES
Volume 21, Issue 7, Pages 1003-1010Publisher
ASIAN-AUSTRALASIAN ASSOC ANIMAL PRODUCTION SOC
DOI: 10.5713/ajas.2008.70392
Keywords
dairy cow; ketosis; liver metabolic characteristic; comparative proteomics; metabolic network
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The objective of the present study was to identify differences in the expression levels of liver proteins between healthy and ketotic cows, establish a liver metabolic interrelationship of ketosis and elucidate the metabolic characteristics of the liver during ketosis. Liver samples from 8 healthy multiparous Hostein cows and 8 ketotic cows were pooled by health status and the proteins were separated by two-dimensional-electrophoresis (2D-E). Statistical analysis of gels was performed using PDQuest software 8.0. The differences in the expression levels of liver proteins (p < 0.05) between ketotic and healthy cows were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF-TOF) tandem mass spectrometry. Five enzyme s/prote ins were identified as being differentially expressed in the livers of ketotic cows: expression of 3-hydroxyacyl-CoA dehydrogenase type-2 (HCDH), acetylcoenzyme A acetyltransferase 2 (ACAT) and elongation factor Tu (EF-Tu) were down-regulated, whereas that of alpha-enolase and creatine kinase were up-regulated. On the basis of this evidence, it could be presumed that the decreased expression of HCDH, which is caused by high concentrations of acetyl-CoA in hepatic cells, in the livers of ketotic cows, implies reduced fatty acid beta-oxidation. The resultant high concentrations of acetyl-CoA and acetoacetyl CoA would depress the level of ACAT and generate more beta-hydroxybutyric acid; high concentrations of acetyl-CoA would also accelerate the Krebs Cycle and produce more ATP, which is stored as phosphocreatine, as a consequence of increased expression of creatine kinase. The low expression level of elongation factor Tu in the livers of ketotic cows indicates decreased levels of protein synthesis due to the limited availability of amino acids, because the most glucogenic amino acids sustain the glyconeogenesis pathway; thus increasing the level of alpha-enolase. Decreased protein synthesis also promotes the conversion of amino acids to oxaloacetate, which drives the Krebs Cycle under conditions of high levels of acetylCoA. It is concluded that the livers of ketotic cows possess high concentrations of acetyl-CoA, which through negative feedback inhibited fatty acid oxidation; show decreased fatty acid oxidation, ketogenesis and protein synthesis; and increased gluconeogenesis and energy production.
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