Seinac Erum P Ctive Ov Roteom Varies P Mics Pro Postpar Ene File And Rtum In Ergy Ba D Warni Dairy C Alance Ing Ass Cows W Sessmen With Neg Nt Of Gative

In recent years,inactive ovaries is a common disease postpartum due to follicle growth abnormalities in the dairy cows,which is characterized by ovarian or related ovarian structure without periodic activity.The growth of ovarian follicles postpartum is affected by a variety of factors in dairy cows.Although negative energy balance is recognized as the main cause of postpartum inactive ovaries,the relationship between protein expression and inactive ovaries postpartum in dairy cows has not still been reported.Therefore,in this experiment the proteomics technology,namely fluorescence two-dimensional differential gel electrophoresis/mass spectrometry(2D-DIGE/MS),combined by bioinformatics analysis were applied to obtain characteristic protein in serum from cows with inactive ovaries postpartum and to clarify the relationship between differential express proteins and inactive ovaries of cows due to negative energy balance postpartum.It will provide the scientific basis for further revealing mechanism of postpartum inactive ovaries of cows with negative energy balance in the future.Test cows,which had similar age and parity in intensive dairy farm,were selected in Heilongjiang Province.The levels of BHBA,NEFA and Glc in serum were measured at 14~21 days postpartum.According to the three parameters,cows were divided into two groups(positive energy balance and negative energy balance).Depend on the estrus behavior and the size of follicular by rectal examination and B ultrasound examination,30 inactive ovaries cows from negative energy balance as experimental group(T)and 30 estrus cows from positive energy balance were selected as control group(C).Serum samples of 15 cows from C or T were detected by 2D-DIGE to obtain proteins profiles and the differential proteins in serum of inactive ovaries cows,and then the relationship between these differential proteins and inactive ovaries of cows from negative energy balance was determined by bioinformatics analysis.The content of AMBP in serum was validated by Western-blot.Another 15 cows from C or T were used to detect serum reproductive hormone and AMBP protein,and correlation between the above indexes was analyzed.Finally,the regression analysis and receiver operating characteristic curve(ROC)were used to determine the AMBP protein as the early warning assessment of inactive ovaries postpartum in dairy cows.Results showed that there are 12 differential protein in serum of cows postpartum with inactive ovaries and estrus cows,including the increase of Apolipoprotein A-I,Apolipoprotein A-IV,Alpha-1-acid glycoprotein,Alpha-2-HS-glycoprotein,Leucine-rich alpha-2-glycop,Complement component 3d(Fragment),Complement C and Endopin 2C,and the decrease of Protein AMBP,Alpha-S1-casein,Hemoglobin subunit beta and Haptoglobin.It was found that these proteins were mainly related to steroid and lipid metabolism,enzyme activity and antioxidant activity according to the bioinformatics analysis.The main pathway involved is fat and vitamin digestion and absorption,PPAR signaling pathway and complement and coagulation cascade.The verified AMBP protein level in serum were consistent with that of 2DDIGE.Serum NEFA levels in T cows were significantly higher than those in C cows at 60~90 days postpartum(p <0.05),serum concentration of E2 was significantly higher than those in C group(p<0.05),serum concentration of AMBP was significantly higher than those in control group(p<0.01).There was significantly positive correlation between the concentration of E2 in serum and the groups(p<0.01).There was a significantly negative correlation between AMBP and the groups(p<0.01).There was significantly positive correlation between serum NEFA and E2(p<0.01).It was determined that the risk of inactive ovaries was increased when AMBP> 4268.59 ng/L in serum by regression analysis and ROC analysis.Conclusion obtained that 2D-DIGE/MS techniques may be used effectively to identify differential proteins in serum of cows postpartum with inactive ovaries due to negative energy balance,and to reveal the relationship between inactive ovaries and differential expression proteins in dairy cows.AMBP may be used to predict risk of inactive ovaries postpartum.It will lay a theoretical and methodological foundation for further studying inactive ovaries postpartum in dairy cows in the future.