Application of quantitative proteomic analysis using tandem mass tags for discovery and identification of novel biomarkers in periodontal disease
Application of quantitative proteomic analysis using tandem mass tags for discovery and identification of novel biomarkers in periodontal disease
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DOI：
10.1002/pmic.201200510
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发表时间：
2013-08-01
期刊：
影响因子：
3.4
通讯作者：
Nomura, Fumio
中科院分区：
文献类型：
--
作者：
Tsuchida, Sachio;Satoh, Mamoru;Nomura, Fumio
牙周病是一种破坏牙龈和口腔周围组织的细菌感染。牙周沟液(GCF)是从牙龈沟和牙袋中提取的。分析牙周液中的生化标志物可以预测牙周病的进展，可能有助于疾病的诊断。然而，目前还没有发现可用于检测牙周病的高灵敏度的GCF生化标志物。因此，寻找牙周病的生化标志物一直是牙周病实验和临床研究的热点。采用串联质量标签(TMT)标记技术，对健康受试者和牙周病患者的牙周组织标本进行分析，通过蛋白质组学分析，共鉴定出619个牙周组织中的GCF蛋白。其中，基质金属蛋白酶(MMP9)和中性粒细胞明胶酶相关脂蛋白(Lcn2)与牙周病的发生发展密切相关。免疫印迹分析显示，牙周病患者牙周组织中MMP9和Lcn2的表达水平明显高于正常对照组。此外，ELISA法还检测到牙周病患者的Lcn2水平明显高于健康人。因此，利用TMT标记对牙周液进行LC-MS/MS分析，可以鉴定出Lcn2，这可能是一种有希望的牙周病检测的牙周液生物标志物。
Periodontal disease is a bacterial infection that destroys the gingiva and surrounding tissues of the oral cavity. Gingival crevicular fluid (GCF) is extracted from the gingival sulcus and pocket. Analysis of biochemical markers in GCF, which predict the progression of periodontal disease, may facilitate disease diagnosis. However, no useful GCF biochemical markers with high sensitivity for detecting periodontal disease have been identified. Thus, the search for biochemical markers of periodontal disease is of continued interest in experimental and clinical periodontal disease research. Using tandem mass tag (TMT) labeling, we analyzed GCF samples from healthy subjects and patients with periodontal disease, and identified a total of 619 GCF proteins based on proteomic analysis. Of these, we focused on two proteins, matrix metalloproteinase (MMP)-9 and neutrophil gelatinase-associated lipocalin (LCN2), which are involved in the progression of periodontal disease. Western blot analysis revealed that the levels of MMP-9 and LCN2 were significantly higher in patients with periodontal disease than in healthy subjects. In addition, ELISA also detected significantly higher levels of LCN2 in patients with periodontal disease than in healthy subjects. Thus, LC-MS/MS analyses of GCF using TMT labeling led to the identification of LCN2, which may be a promising GCF biomarker for the detection of periodontal disease.