荧光双向电泳检测雌雄小鼠肝组织蛋白组学的差异

发布时间：2018-01-07 22:09

本文关键词：荧光双向电泳检测雌雄小鼠肝组织蛋白组学的差异　出处：《中国比较医学杂志》2017年10期 　论文类型：期刊论文

【摘要】：目的检测雌雄小鼠肝组织蛋白质组表达差异,探讨肝病发生的性别差异机制。方法采用双向荧光差异凝胶电泳(two dimension difference gel electrophoresis,2D-DIGE)和基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF-MS)技术检测C57BL/6J雌雄小鼠肝组织的蛋白质组差异表达谱,分离并鉴定差异表达蛋白,应用Western blot进行验证,并对差异表达蛋白进行生物信息学分析,包括蛋白功能注释、分类分析、京都基因与基因组百科全书通路分析。结果经双向荧光差异凝胶电泳-图像分析得到1767个蛋白点,其中差异倍数≥1.5(P0.05)的蛋白点325个。从中选择78个差异蛋白点进行MALDI-TOF-MS鉴定,得到48种差异性表达的蛋白质。其中,与雌鼠相比,雄鼠肝组织中高表达的蛋白有14种,低表达的蛋白有34种。选取6个差异蛋白点进行Western blot验证,证明了2D-DIGE结果的可靠性。GO分析结果显示,涉及雌雄鼠肝组织中的差异蛋白在细胞组分、分子功能、生物学过程中的分布广泛。KEGG通路分析发现差异蛋白涉及6条信号通路。结论 C57BL/6J雌雄小鼠肝组织的差异性蛋白表达的检测结果为研究性别差异性肝病发生的分子机制提供了基础生物学信息和有益的线索。
[Abstract]:Objective to detect the expression difference between male and female mice liver tissue proteome, explore the mechanism of gender differences in liver disease. Methods using two-dimensional difference gel electrophoresis (two dimension difference gel electrophoresis, 2D-DIGE) and matrix assisted laser desorption ionization time-of-flight mass spectrometry (matrix-assisted laser desorption/ionization time of flight mass spectrometry, MALDI-TOF-MS) differential expression of proteome in detecting C57BL/6J male and female mouse liver tissue, the expression of protein isolate and identify the differences in application of Western blot to verify, and the differentially expressed protein by bioinformatics analysis, including protein functional annotation, classification analysis, analysis of Kyoto Encyclopedia of genes and genomes pathway. Results by fluorescence two-dimensional difference gel electrophoresis image analysis of 1767 protein spots, among them the difference is more than 1.5 times (P0.05) 325 protein spots. Choose from MALDI-TOF-MS identified 78 differential protein spots, 48 proteins were differentially expressed. Among them, compared with female mice, the high expression of liver tissue in male rats there are 14 kinds of proteins, low expression of protein 34. Selected 6 differentially expressed protein spots were Western blot validation,.GO reliability analysis results show that 2D-DIGE the results show that relates to differences in male and female rats in liver tissue protein in the cellular component, molecular function, biological process in the widely distributed.KEGG pathway analysis showed that the differential proteins involved in 6 pathways. Conclusion C57BL/6J male and female mouse liver tissue to detect differentially expressed proteins occurred for the study of gender differences in the molecular mechanism of liver disease the basic biological information and useful clues.

【作者单位】：大连医科大学实验动物中心;
【基金】：辽宁省教育厅科学规划课题(L2014341) 国家自然科学基金(30872950)
【分类号】：R-33;R575
【正文快照】： confirmed the reliability of 2D-DIGE results.GO analysis showed that the differentially expressed proteins in the liver tissuesof male and female mice are associated to various cellular component,molecular function and biological process.6pathways were s

【相似文献】