ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2014, Vol. 45 ›› Issue (5): 844-852.doi: 10.11843/j.issn.0366-6964.2014.05.025

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Screening Protein Markers in Urine of Dairy Cows with Fatty Liver Disease Based on ITRAQ-HPLC-MS Technology

SUN Yu-hang, XIA Cheng*, SHU Shi, SUN Ling-wei, XU Chuang   

  1. (College of Animal Science and Veterinary Medicine,Heilongjiang Bayi Agricultural University,Daqing 163319, China)
  • Received:2013-10-30 Online:2014-05-23 Published:2014-05-23

Abstract:

To obtain new protein markers of fatty liver disease in dairy cows,the relative and absolute quantification isotope labeling (iTRAQ) technology was used to screen and identify differentially expressed proteins in urine of dairy cows with fatty liver disease.40 cows urines were collected and divided into diseased groups A1,A2 and healthy groups B1,B2,respectively including 10 cows in each group.Every 10 urine samples in the same group was equally mixed into 1 sample for the experiment.Subsequently,the samples which the requirements were marked by an iTRAQ kit.Finally,the high-performance liquid chromatography (HPLC) and tandem mass spectrometry were used.In this way,we got a series of data as a result,then the results were conducted by a series of bioinformatics analysis.Totally,110 differentially expressed proteins were screened,in which 50 were upregulated and 60 were downregulated.4 key proteins were acquired by bioinformatic analysis:Vitronectin (VTN),Lipocalin (LCN2),Prothrombin (F2) and Clusterin (CLU).Differentially expressed proteins in the urine of dairy cows with fatty liver disease were screened in this experiment,4 key regulatory factors were closely related to fatty liver disease,and should be further confirmed as important biological markers of fatty liver disease in dairy cows,which will provide a new method in early detection and diagnosis of fatty liver disease of dairy cows.

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