不同生长时期梅花鹿鹿茸差异蛋白质组学分析
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- 英文论文题名：Comparative Proteomic Analysis in Different Growth Stages of Sika Deer Velvet Antler
- 论文作者：张然然 ; 刘华淼 ; 邵元臣 ; 周盼伊 ; 苏莹 ; 王磊 ; 邢秀梅
- 英文论文作者：Zhang Ranran ; Liu Huamiao ; Shao Yuanchen ; Zhou Panyi ; Su Ying ; Wang Lei ; Xing Xiumei ; State Key Laboratory of Special Economic Animal Molecular Biology ; Institute of Special Wild Economic Animal and Plant Science ; Chinese Academy Agricultural Sciences
- 年：2016
- 作者机构：中国农业科学院特产研究所特种经济动物分子生物重点实验室;
- 论文关键词：梅花鹿 ; 鹿茸 ; 不同生长时期 ; 双向凝胶电泳 ; 蛋白质组学
- 英文论文关键词：sika deer ; velvet antler ; different growth stages ; two dimensional gel electrophoresis ; proteome
- 会议召开时间：2016-06-01
- 会议录名称：“十二五” 鹿科学研究进展
- 语种：中文
- 分类号：S825
- 学会代码：ZXMY
- 学会名称：中国畜牧业协会
- 页数：13
- 文件大小：1228k
- 原文格式：O
摘要
为了解梅花鹿鹿茸生长过程中蛋白质表达特征,从分子水平认识鹿茸生长机制,本研究以10d、40d、60d与130d的梅花鹿鹿茸为试验材料,运用双向凝胶电泳(2-DE)和MALDI-TOF/TOF质谱鉴定技术对梅花鹿鹿茸生长过程中差异表达蛋白质进行筛选,并结合生物信息学方法对差异表达蛋白质进行功能分析。结果显示有46种蛋白质差异性表达,且主要参与了细胞骨架、转运过程、信号转导、细胞凋亡、骨发育、蛋白质合成、核酸代谢、免疫、能量代谢、细胞增殖、抗氧化、蛋白质折叠等生物学过程。结合鹿茸的快速生长与快速骨化的独特生长过程,对差异表达蛋白质中的骨发育相关蛋白、抗氧化蛋白、细胞凋亡相关蛋白做进一步分析,发现P4HB、SPARC、过氧化物还原酶2、过氧化物还原酶4、半乳糖凝集素1、视黄酸结合蛋白1等6种蛋白质在鹿茸快速生长与快速骨化过程中起着重要的作用,为鹿茸生长与骨化机制的进一步研究奠定基础。
To investigate into the proteome profiling of the velvet antler and clarify velvet antler development mechanism in the level molecular biology,the sika deer velvet antlers aged10 d、40 d、60 d and 130 d were selected as experimental material.Two-dimensional gel electrophoresis(2- DE) was employed to separate and quantify proteins of the velvet antlers during their four days development,the differentially expressed proteins were identified by MALDI- TOF /TOF- MS,and the function of proteins were analyzed using bioinformatics methods.The result showed that 46 proteins were differentially expressed significantly,which were involved the biology process of cytoskeleton、transportation、signal transduction、apoptotic process、bone development、protein synthesis、nucleotide biosynthesis、immunity、energy metabolism、cell proliferation and antioxidant activity.We focused on the proteins involved in bone development、antioxidant activity and apoptotic process due to the special growth process with rapid growth and ossification,and found that P4HB、SPARC、peroxiredoxin 2、peroxiredoxin 4、galectin 1 and retinoic acid binding protein 1 may play vital role in velvet antler growth and ossification process.Taken together,the result provided a basis for further research on the molecular mechanisms involved in the accelerated growth and ossification of deer velvet antler.
To investigate into the proteome profiling of the velvet antler and clarify velvet antler development mechanism in the level molecular biology,the sika deer velvet antlers aged10 d、40 d、60 d and 130 d were selected as experimental material.Two-dimensional gel electrophoresis(2- DE) was employed to separate and quantify proteins of the velvet antlers during their four days development,the differentially expressed proteins were identified by MALDI- TOF /TOF- MS,and the function of proteins were analyzed using bioinformatics methods.The result showed that 46 proteins were differentially expressed significantly,which were involved the biology process of cytoskeleton、transportation、signal transduction、apoptotic process、bone development、protein synthesis、nucleotide biosynthesis、immunity、energy metabolism、cell proliferation and antioxidant activity.We focused on the proteins involved in bone development、antioxidant activity and apoptotic process due to the special growth process with rapid growth and ossification,and found that P4HB、SPARC、peroxiredoxin 2、peroxiredoxin 4、galectin 1 and retinoic acid binding protein 1 may play vital role in velvet antler growth and ossification process.Taken together,the result provided a basis for further research on the molecular mechanisms involved in the accelerated growth and ossification of deer velvet antler.
引文
[1]SUI Z G,ZHANG L H,HUO Y S,et al.Bioaetive components of velvet antlers and their pharmacological properties[J].J Pharm Biomed Anal,2014,87;229-240.
[2]PARK H J,LEE D H,PARK S G.Proteome analysis of red deer antlers[J].Proteomics,2004,4(11):3642-3653.
[3]BRADFORD M M.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J].Anal Biochem,1976,72(1):248-254.
[4]LI C,SUTTIE J M.Light microscopic studies of pedicle and early first antler development in red deer(Cervus elaphus)[J].Anat Rec,1994,239(2):198-215.
[5]CHUA J,SEET L F,JIANG Y.Increased SPARC expression in primary angle closure glaucoma iris[J].Mol Vis,2008,14:1 886-1 892.
[6]PAJUNEN L,JONES T A,GODDARD A.Regional assignment of the human gene coding for a multifunctional polypeptide(P4HB)acting as the beta-subunit of prolyl 4-hydroxylase and the enzyme protein disulfide isomerase to 17q25[J].Cytogenet Cell Genet,1991,56(3-4):165-168.
[7]TERMINE J D,KLEINMAN H K,WHITSON S W,et al.Osteonectin,a bone-specific protein linking mineral to collagen[J].Cell,1981,26(1):99-105.
[8]BRADSHAW A D,SAGE E H.SPARC,a matricellular protein that functions in cellular differentiation and tissue response to injury[J].J Clin Invest,2001,107(9):1 049-1 054.
[9]DELANY A M,HANKENSON K D.Thrombospondin-2 and SPARC/osteonectin are critical regulators of bone remodeling[J].J Cell Commun Signal,2009,3(3—4):227-238.
[10]REIS L M D,KESSLER C B,ADAMS D J,et al.Accentuated osteoclastic response to parathyroid hormone undermines bone mass acquisition in osteonectin-null mice[J].Bone,2008,43(2):264-273.
[11]GIUSTARINI D,DALLE-DONNE I,TSIKAS D,et al.Oxidative stress and human diseases:Origin,link,measurement,mechanisms,and biomarkers[J].Crit Rev Clin Lab Sci,2009,46(5-6):241-281.
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[13]赵东.梅花鹿鹿茸双向电泳体系建立及蛋白质组学的研究[D].镇江:江苏科技大学,2012.ZHAO D.Establishment of two-dimensional electrophoresis system and proteome analysis of Sika deer antler.Zhenjiang;Jiangsu University of Science and Technology,2012.
[14]LI C,HARPER A,PUDDICK J,et al.Proteomes and signalling pathways of antler stem cells[J].PLoSOne,2012,7(1):e30026.
[15]MARTIN A,S F D,EDUARDO P,et al.Typical 2-Cys peroxiredoxins--modulation by covalent transformations and none ovalent interactions[J].FEBS Journal(Online),2009,276(9):2 478-2 493.
[16]RHEE S G,CHAE H Z,KIM K.Peroxiredoxins;A historical overview and speculative preview of novel mechanisms and emerging concepts in cell signaling[J].Free Radical Bio Med,2005,38(12):1 543-1 552.
[17]任丽平.鮸鱼过氧化物还原酶Peroxiredoxin基因的分子克隆与表达分析[D].浙江:浙江海洋学院.2014.REN L P.Molecular cloning and expression of peroxiredoxins from Miichthys miiuy[D].Zhejiang;Zhejiang Ocean University,2014.
[18]冯海华,闭兴明,赵丽红.胰岛素样生长因子1对不同生长时期鹿茸生长中心细胞体外增殖的影响[J].中国组织工程研究与临床康复,2007(37):7 373-7 376.FENG H H,BI X M,ZHAO L H.Effects of insulin-like growth factor 1 on the in vitro proliferation of antler organic center cells at different growing periods[J].Journal of Clinical Rehabilitative Tissue Engineering Research,2007(37):7 373-7 376.
[19]LU Y,LOTAN D,LOTAN R.Differential regulation of constitutive and retinoic acid-induced galectin-1 gene transcription in murine embryonal carcinoma and myoblastic cells[J].Biochim Biophys Acta,2000,1491(1-3):13-19.
[20]BLUM N,BEGEMANN G.The roles of endogenous retinoid signaling in organ and appendage regeneration[J].Cell Mol Life Sci,2013,70(20):3907-3927.
[21]WESTON A D,HOFFMAN L M,UNDERHILL T M.Revisiting the role of retinoid signaling in skeletal development[J].Birth Defects Res C Embryo Today,2003,69(2):156-173.
[2]PARK H J,LEE D H,PARK S G.Proteome analysis of red deer antlers[J].Proteomics,2004,4(11):3642-3653.
[3]BRADFORD M M.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J].Anal Biochem,1976,72(1):248-254.
[4]LI C,SUTTIE J M.Light microscopic studies of pedicle and early first antler development in red deer(Cervus elaphus)[J].Anat Rec,1994,239(2):198-215.
[5]CHUA J,SEET L F,JIANG Y.Increased SPARC expression in primary angle closure glaucoma iris[J].Mol Vis,2008,14:1 886-1 892.
[6]PAJUNEN L,JONES T A,GODDARD A.Regional assignment of the human gene coding for a multifunctional polypeptide(P4HB)acting as the beta-subunit of prolyl 4-hydroxylase and the enzyme protein disulfide isomerase to 17q25[J].Cytogenet Cell Genet,1991,56(3-4):165-168.
[7]TERMINE J D,KLEINMAN H K,WHITSON S W,et al.Osteonectin,a bone-specific protein linking mineral to collagen[J].Cell,1981,26(1):99-105.
[8]BRADSHAW A D,SAGE E H.SPARC,a matricellular protein that functions in cellular differentiation and tissue response to injury[J].J Clin Invest,2001,107(9):1 049-1 054.
[9]DELANY A M,HANKENSON K D.Thrombospondin-2 and SPARC/osteonectin are critical regulators of bone remodeling[J].J Cell Commun Signal,2009,3(3—4):227-238.
[10]REIS L M D,KESSLER C B,ADAMS D J,et al.Accentuated osteoclastic response to parathyroid hormone undermines bone mass acquisition in osteonectin-null mice[J].Bone,2008,43(2):264-273.
[11]GIUSTARINI D,DALLE-DONNE I,TSIKAS D,et al.Oxidative stress and human diseases:Origin,link,measurement,mechanisms,and biomarkers[J].Crit Rev Clin Lab Sci,2009,46(5-6):241-281.
[12]徐代勋.梅花鹿鹿茸角柄骨膜不同部位差异蛋白的筛选[D].镇江:江苏科技大学,2011.XU D X.Screening of differential proteins in different parts of pedicle periosteum of Sika deer.Zhenjiang;Jiangsu University of Science and Technology,2011.
[13]赵东.梅花鹿鹿茸双向电泳体系建立及蛋白质组学的研究[D].镇江:江苏科技大学,2012.ZHAO D.Establishment of two-dimensional electrophoresis system and proteome analysis of Sika deer antler.Zhenjiang;Jiangsu University of Science and Technology,2012.
[14]LI C,HARPER A,PUDDICK J,et al.Proteomes and signalling pathways of antler stem cells[J].PLoSOne,2012,7(1):e30026.
[15]MARTIN A,S F D,EDUARDO P,et al.Typical 2-Cys peroxiredoxins--modulation by covalent transformations and none ovalent interactions[J].FEBS Journal(Online),2009,276(9):2 478-2 493.
[16]RHEE S G,CHAE H Z,KIM K.Peroxiredoxins;A historical overview and speculative preview of novel mechanisms and emerging concepts in cell signaling[J].Free Radical Bio Med,2005,38(12):1 543-1 552.
[17]任丽平.鮸鱼过氧化物还原酶Peroxiredoxin基因的分子克隆与表达分析[D].浙江:浙江海洋学院.2014.REN L P.Molecular cloning and expression of peroxiredoxins from Miichthys miiuy[D].Zhejiang;Zhejiang Ocean University,2014.
[18]冯海华,闭兴明,赵丽红.胰岛素样生长因子1对不同生长时期鹿茸生长中心细胞体外增殖的影响[J].中国组织工程研究与临床康复,2007(37):7 373-7 376.FENG H H,BI X M,ZHAO L H.Effects of insulin-like growth factor 1 on the in vitro proliferation of antler organic center cells at different growing periods[J].Journal of Clinical Rehabilitative Tissue Engineering Research,2007(37):7 373-7 376.
[19]LU Y,LOTAN D,LOTAN R.Differential regulation of constitutive and retinoic acid-induced galectin-1 gene transcription in murine embryonal carcinoma and myoblastic cells[J].Biochim Biophys Acta,2000,1491(1-3):13-19.
[20]BLUM N,BEGEMANN G.The roles of endogenous retinoid signaling in organ and appendage regeneration[J].Cell Mol Life Sci,2013,70(20):3907-3927.
[21]WESTON A D,HOFFMAN L M,UNDERHILL T M.Revisiting the role of retinoid signaling in skeletal development[J].Birth Defects Res C Embryo Today,2003,69(2):156-173.