目的应用定量蛋白质组学技术筛选周期性机械应力条件下大鼠软骨细胞差异表达蛋白质。方法将体外培养的大鼠软骨细胞随机分为对照组和周期性机械应力组,分别提取细胞蛋白,并依次进行同位素标记、液相色谱分离、质谱鉴定和生物信息学分析。结果共鉴定出5468个蛋白,差异性表达蛋白为485个;其中上调168个,下调317个。经生物信息学分析,共有478个蛋白被6079条基因本体论(GO)功能条目注释,230个蛋白参与了241条京都基因与基因组百科全书(KEGG)信号通路。结论成功构建周期性机械应力条件下大鼠软骨细胞差异蛋白质表达谱,为周期性机械应力促进大鼠软骨细胞增殖和细胞外基质合成的机制研究提供实验基础。
Objective To screen the differentially expressed proteins of rat chondrocytes under periodic mechanical stress by quantitative proteomics technique.Methods Rat chondrocytes were cultured in vitro and then divided into two groups of A(normal control)and B(under periodic mechanical stress).The total proteins in groups of A and B were extracted and labeled by isobaric tags for relative and absolute quantitation,respectively.Then the labeled proteins were isolated by liquid chromatography,identified by Q-exactive mass spectrometry and analyzed by bioinformatics for comparing the differentially expressed proteins.Results A total of 5468 proteins was identified.The differentially expressed proteins were 485,among which 168 proteins were upregulated and 317 proteins were downregulated.The bioinformatics analysis indicated that 478 proteins were involved in6079 gene ontology(GO)function entries and 230 proteins were participated in 241 kyoto encyclopedia of genes and genomes(KEGG)signal pathways.Conclusion The differentially expressed proteome profile of rat chondrocytes under periodic mechanical stress has been successfully constructed,which may provide a foundation for studying the underlying mechanism of proliferation of rat chondrocytes and synthesis of extracellular matrix promoted by periodic mechanical stress.