摘要
目的应用组织蛋白质组学方法寻找与成人股骨头缺血性坏死(ONFH)相关的蛋白质。方法于非创伤性ONFH患者股骨头中分别采集坏死骨组织样品与正常骨组织样品各12份,采用四步溶剂法提取骨组织中的总蛋白,应用多维液相色谱与串联质谱联用技术和生物信息学方法对组织蛋白进行分离鉴定和功能蛋白质组学分析。结果坏死组织样本和正常组织样本中鉴定二肽段以上的高可信度蛋白质数量分别为1302个和99个,假阳性率为0·9%。通过与对照组的质谱计数,在坏死骨组织中鉴定了141个高表达蛋白和56个低表达蛋白。基因本体论蛋白功能分析发现34种蛋白质与ONFH密切相关。本实验在股骨头坏死组织中证实ChST2基因(硫酸软骨素蛋白聚糖合成过程中的关键蛋白)和GPCR26基因(介导细胞内钙离子动员的蛋白质)的低表达。结论非创伤性ONFH存在复杂的信号传导通路。硫酸软骨素蛋白聚糖的生物合成和多种离子的转运、动员可能参与ONFH发病机制中的关键过程。研究发现的差异表达蛋白可能成为ONFH早期临床诊断的候选生物学标志物。
Objective To explore proteins associated with non-traumatic osteonecrosis of the femoral head （ONFH） in adults by high throughput proteomic approach. Methods 12 samples harvested respectively from the femoral head of the patients with ONFH and those of fresh femoral neck fracture were included respectirely in necrotic group and control group. Four-step sequential extraction was used to collect total proteins from the bone tissue. Multi-dimensional liquid chromatography-tandem mass spectrometry was applied to separate and identify the extracted proteins, and bioinformatics analysis was carried out to accomplish functional proteomic research. Results The proteome analysis of bone tissue in necrotic group and control group revealed that the high-confidence proteins based on two peptides minimum were 1302 and 999 respectively, with a false-positive rate of 0. 9%. Using spectral counting, we identified 141 overexpressed and 56 underexpressed proteins comparing ONFH bones with the controls. Enrichment analysis in Gene Ontology categories of the differentially expressed gene found that 34 differential expressed proteins were highly associated with osteonecrosis of the femoral head. The underexpression of ChST2, a key protein involved in biosynthesis of chondroitin sulfate proteoglycans, and the underexpression of GPCR26, a protein that mediates intracellular calcium mobilization, were verified in ONFH bones. Conclusions Our research revealed that ONFH is associated with complicated signal transduction pathway, and that biosynthesis of chondroitin sulfate proteoglycans and cation transport and mobilization may be a key process involved in the pathogenesis of ONFH. The identified differential expressed proteins might be promising biomarker candidates for early diagnosis of the disease.
出处
《中华关节外科杂志（电子版）》
CAS
2011年第4期35-40,共6页
Chinese Journal of Joint Surgery(Electronic Edition)
关键词
股骨头坏死
蛋白质组学
色谱法
液相
光谱分析
质量
信号传导
Femur head necrosis
Proteomics
Chromatography, liquid
Spectrum analysis, mass
Signal transduction
