荷斯坦奶牛产后期血浆差异蛋白质组学研究
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摘要
为了检测和分析荷斯坦奶牛分娩后不同时期血浆中的差异蛋白。采用二维凝胶电泳(2-DE)分离血浆蛋白,考马斯亮蓝染色,PDQuest8.0软件检测分析差异蛋白点,MADIL-TOF-TOF串联质谱鉴定差异蛋白点。结果发现3种差异蛋白,分别是结合珠蛋白、免疫球蛋白γ2链C区和3,5-二碘水杨酸络合的牛血清白蛋白。对3种蛋白在产后不同时期的趋势进行分析,可能分别与产后应激、免疫应答和物质运输有关。荷斯坦奶牛产后不同时期血浆中蛋白表达存在差异,对差异表达蛋白的研究有利于探索奶牛产后生理机制,可作为产后饲养、配种及防止产后感染等研究的基础。
Proteome in plasma from Holstein cows in the different postpartum periods was analyzed in this research.Two-dimensional gel electrophoresis(2-DE)was used to separate plasma protein and stained with colloidal coomassie blue,differential expression proteins were detected by PDQuest 8.0software and were identified by MADIL-TOF-TOF tandem mass spectrometry.Three differential proteins were found,including haptoglobin,Ig gamma-2 chain C region(clone 32.2)-bovine(fragment),bovine serum albumin in complex with 3,5-diiodosalicylic acid.The content variation tendency of the three proteins in different periods were analyzed.Three proteins may be associated with postpartum stress,immune response and material transportation,respectively.There were significant differences in protein levels of plasma from Holstein cows in the different postpartum periods,which may be helpful to explore cows postpartum physiological mechanism,and can be used as the basis of the research on postnatal feeding,breeding and preventing from postpartum infection.
Proteome in plasma from Holstein cows in the different postpartum periods was analyzed in this research.Two-dimensional gel electrophoresis(2-DE)was used to separate plasma protein and stained with colloidal coomassie blue,differential expression proteins were detected by PDQuest 8.0software and were identified by MADIL-TOF-TOF tandem mass spectrometry.Three differential proteins were found,including haptoglobin,Ig gamma-2 chain C region(clone 32.2)-bovine(fragment),bovine serum albumin in complex with 3,5-diiodosalicylic acid.The content variation tendency of the three proteins in different periods were analyzed.Three proteins may be associated with postpartum stress,immune response and material transportation,respectively.There were significant differences in protein levels of plasma from Holstein cows in the different postpartum periods,which may be helpful to explore cows postpartum physiological mechanism,and can be used as the basis of the research on postnatal feeding,breeding and preventing from postpartum infection.
引文
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[17]MORETTI P,PROBO M,MORANDI N,et al.Early post-partum hematological changes in Holstein dairy cows with retained placenta[J].Anim Reprod Sci,2015,152:17-25.
[18]HUZZEY J,DUFFIELD T,LEBLANC S,et al.Short communication:Haptoglobin as an early indicator of metritis[J].J Dairy Sci,2009,92(2):621-625.
[19]WAI-HOE L,WING-SENG L,ISMAIL Z,et al.Proteomics and detection of uromodulin in first-time renal calculi patients and recurrent renal calculi patients[J].Appl Biochem Biotech,2009,159(1):221-232.
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[21]王媛,刘春艳,赵久阳.乙型肝炎病毒相关性肾炎的尿液蛋白质谱研究[J].中华生物医学工程杂志,2013,19(4):300-305.
[22]陈昌辉,叶长宁.母婴血清IgG及亚类含量与胎盘转运率[J].中国优生优育,2001,12(4):157-159.
[2]MAEDA Y,OHTSUKA HOIKAWA M.Effect of the periparturient period on blood free amino acid concentration in dairy cows/healthy cows[J].J Vet Med Anim Health,2012,4(9):124-129.
[3]王聪,刘强,张延利,等.丙二醇对奶牛围产期血浆激素和肝脏糖异生限速酶的影响[J].核农学报,2013,27(1):132-137.
[4]冯番,甄二英,许睛.奶牛围产期营养与产后病[J].饲料博览(技术版),2007(4):36-37.
[5]叶锋.围产期产犊应激,代谢障碍及免疫力变化对奶牛的影响及应对措施[J].中国乳业,2010(12):32-32.
[6]杨永新,王加启,卜登攀,等.基于2-DE结合质谱研究围产期奶牛血浆蛋白表达的变化[J].畜牧兽医学报,2010,41(2):188-192.
[7]STEEL L F,SHUMPERT D,TROTTER M,et al.A strategy for the comparative analysis of serum proteomes for the discovery of biomarkers for hepatocellular carcinoma[J].Proteomics,2003,3(5):601-609.
[8]ASCHBACHER P,FEIL V.Metabolism of 3,5-diiodosalicylic acid in cattle and rats[J].J Dairy Sci,1968,51(5):762-766.
[9]ASCHBACHER P,CRAGLE R,SWANSON E,et al.Metabolism of oral iodide and 3,5-diiodosalicylic acid in the pregnant cow[J].J Dairy Sci,1966,49(9):1042-1045.
[10]LEE J,KING W A,CONLEY C.Protein-bound iodine in the blood of dairy cattle fed a trace-mineralized salt[J].J Dairy Sci,1960,43(4):512-518.
[11]牛丽莉,刘涛,魏彩虹,等.分娩前后奶牛血浆差异表达蛋白初步分析[J].畜牧兽医学报,2010,41(9):1090-1094.
[12]KHAZAEI H A,TEYMURI B,NAKHAEI A,et al.Association of haptoglobin phenotypes with clinical features of preterm labor disease[J].Acta Medica Iranica,2013,51(8):554-559.
[13]CHO J K,KIM Y H,PARK I Y,et al.Polymorphism of haptoglobin in patients with premature rupture of membrane[J].Yonsei Med J,2009,50(1):132-136.
[14]YANG Y,WANG J,BU D,et al.Comparative proteomics analysis of plasma proteins during the transition period in dairy cows with or without subclinical mastitis after calving[J].Czech J Anim Sci,2012,57:481-489.
[15]SABEDRA D A.Serum haptoglobin as an indicator for calving difficulties and postpartal diseases in transition dairy cows[J].Dairy Cows,2012.
[16]LARSSON A,PALM M,HANSSON L O,et al.Reference values forα1-acid glycoprotein,α1-antitrypsin,albumin,haptoglobin,C-reactive protein,IgA,IgG and IgM during pregnancy[J].Acta Obstet Gyn Scan,2008,87(10):1084-1088.
[17]MORETTI P,PROBO M,MORANDI N,et al.Early post-partum hematological changes in Holstein dairy cows with retained placenta[J].Anim Reprod Sci,2015,152:17-25.
[18]HUZZEY J,DUFFIELD T,LEBLANC S,et al.Short communication:Haptoglobin as an early indicator of metritis[J].J Dairy Sci,2009,92(2):621-625.
[19]WAI-HOE L,WING-SENG L,ISMAIL Z,et al.Proteomics and detection of uromodulin in first-time renal calculi patients and recurrent renal calculi patients[J].Appl Biochem Biotech,2009,159(1):221-232.
[20]CHO W,JUNG K,REGNIER F E.Sialylated Lewis x antigen bearing glycoproteins in human plasma[J].J Proteome Res,2010,9(11):5960-5968.
[21]王媛,刘春艳,赵久阳.乙型肝炎病毒相关性肾炎的尿液蛋白质谱研究[J].中华生物医学工程杂志,2013,19(4):300-305.
[22]陈昌辉,叶长宁.母婴血清IgG及亚类含量与胎盘转运率[J].中国优生优育,2001,12(4):157-159.