梅花鹿致敏与休眠鹿茸干细胞差异蛋白表达的2D-DIGE分析

doi:10.11843/j.issn.0366-6964.2016.01.013

摘要/Abstract

摘要：

旨在对梅花鹿(Cervus nippon)致敏鹿茸干细胞与休眠鹿茸干细胞表达蛋白进行差异筛选、鉴定及生物信息分析，为深入探讨鹿茸独特的再生分子调节机制奠定基础。本研究采用双向荧光差异凝胶电泳(Two-dimensional fluorescence difference in gel electrophoresis，2D-DIGE)分离蛋白样品；利用DeCyder 7.2 分析软件对2D-DIGE图像进行统计学分析寻找差异表达蛋白；利用MALDI-TOF-MS (Matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry)鉴定差异蛋白，通过Mascot 软件搜索NCBInr 数据库寻找匹配的蛋白；采用PANTHER (Protein Analysis Through Evolutionary Relationships) 软件对差异蛋白进行聚类分析，REACTOME数据库分析差异蛋白所参与的信号通路。结果得到了致敏鹿茸干细胞与休眠鹿茸干细胞2D-DIGE图谱，致敏鹿茸干细胞与休眠鹿茸干细胞蛋白丰度相比较，比值≥1.1倍以及比值≤-1.1倍(P＜0.05)的差异蛋白点有159个，其中110个上调表达，49个下调表达，EDA(Extended data analysis)分析得到了多个Marker蛋白，质谱鉴定了84 个差异蛋白质点，48个为阳性结果，共来自27 种蛋白质。并对已鉴定蛋白进行了GO分析以及信号通路富集分析。致敏鹿茸干细胞与休眠鹿茸干细胞蛋白差异明显，质谱鉴定获得了来自多种可能与鹿茸再生相关的差异蛋白。由此可知，鹿茸再生是鹿茸干细胞从休眠到致敏的转化过程，需要多种蛋白分子以及信号通路的综合调控。

关键词: 鹿茸干细胞, 再生, 蛋白质组学, 2D-DIGE

Abstract:

The objective of this study was to screen，identify and analyze the differentially expressed proteins in the potentiated and dormant antler stem cells in sika deer (Cervus nippon)，and then to shed lights on the molecular mechanisms underlying antler regeneration.The two-dimensional fluorescence of gel electrophoresis (2D-DIGE) was used to separate the protein spots；Differentially expressed protein spots were selected by DeCyder 2D (version 7.2)；Matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF-MS) was carried out to obtain peptide mass fingerprinting，Mascot software was used to search the matched proteins in the NCBInr database；PANTHER (Protein Analysis Through Evolutionary Relationships) and REACTOME analysis were performed to further explore the involved signal pathways about these identified proteins.The proteomic profile of the potentiated antler stem cells compared with the dormant antler stem cells was explored by 2D-DIGE.One hundred fifty-nine protein spots with more than 1.1-fold changes and less than -1.1-fold changes and P values less than 0.05 differentially expressed by the potentiated over the dormant antler stem cells，including 110 up-regulated and 49 down-regulated protein spots.Multiple markers were obtained by extended data analysis(EDA) module.MALDI-TOF-MS identified 84 differentially expressed protein spots and 48 of them which came from 27 kinds of proteins were positive.There is a significant difference at proteomic level between the potentiated and the dormant antler stem cells，and some identified proteins which are involved in multiple functional categories might be related to antler regeneration.Therefore，antler regeneration is a process from the dormant to the potentiated states in antler stem cells，which is regulated by multiple proteins and a complicated signal network.

Key words: antler stem cell, regeneration, proteomics, 2D-DIGE

中图分类号:

S825
S813.3

董振，王权威，刘振，孙红梅，李春义. 梅花鹿致敏与休眠鹿茸干细胞差异蛋白表达的2D-DIGE分析[J]. 畜牧兽医学报, 2016, 47(1): 91-104.

DONG Zhen，WANG Quan-wei，LIU Zhen，SUN Hong-mei，LI Chun-yi. Analysis of Differentially Expressed Proteins in the Potentiated and Dormant Antler Stem Cells through 2D-DIGE[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(1): 91-104.

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