牙周炎与慢性阻塞性肺疾病患者的唾液多肽组学分析

王蕾; 李雪芬; 张天翼; 李威; 王左敏; 林梅

doi:10.12182/20230160505

牙周炎与慢性阻塞性肺疾病患者的唾液多肽组学分析

Salivary Peptide Profiling Analysis of Patients with Periodontitis and Chronic Obstructive Pulmonary Disease

摘要

摘要:
目的分析牙周炎（periodontitis, PD）与慢性阻塞性肺疾病（chronic obstructive pulmonary disease, COPD）患者唾液中与疾病相关的差异性表达多肽，寻找具有潜在诊断意义的生物标志物，为探讨COPD的早期防治开辟新视角。
方法筛选PD患者10例（PD组）、PD伴COPD患者10例（PD伴COPD组）及健康对照人群8例（对照组），收集各组临床资料及唾液样本。应用弱阳离子交换磁珠（weak-cation-exchange magnetic beads, WCX-MB）分离纯化唾液上清样本，应用基质辅助激光解析电离飞行时间质谱（matrix-assisted laser desorption ionization-time of flight mass spectrometry, MALDI-TOF-MS）系统获取样本生物学信息，进行多肽差异性质谱分析，筛选出组间差异性多肽，并通过液相色谱-质谱/质谱联用技术（liquid chromatography tandem mass spectrometry, LC-MS/MS）对差异性多肽进行鉴定。
结果三组样本共检测出77个差异多肽峰，其中PD伴COPD组和PD组间显著性差异多肽峰10个，PD伴COPD组有8个多肽峰（1193.5、1836.2、1735.1、1321.3、1356.8、2086.8、1863.6、2230.9）表达增高，2个多肽峰（1067.3、1124.4）表达减低。其中1193.5、1356.8多肽峰被鉴定为富组蛋白1、颌下腺激素调节蛋白3B及唾液酸性富脯磷蛋白1/2。
结论本研究通过WCX-MB和MALDI-TOF-MS分析出PD伴COPD患者唾液中存在与疾病相关的差异性表达多肽，筛选出的差异性表达多肽有望成为早期诊断COPD的辅助指标。

Abstract:
Objective To analyze the salivary peptide profiles of patients with periodontitis (PD) and chronic obstructive pulmonary disease (COPD), to identify differentially expressed peptides that are associated with diseases, to explore for biomarkers with potential diagnostic significance, and to probe for new perspectives for the early prevention and treatment of COPD.
Methods A total of 10 PD patients (the PD group), 10 PD patients with COPD (the PD plus COPD group), and 8 healthy controls (the Control group) were selected for the study. The clinical data and saliva samples of the subjects were collected. Salivary supernatant samples were separated and purified with weak-cation-exchange magnetic bead-based (WCX-MB). With matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS), the biodata of the samples were obtained and differential salivary peptide profiling was conducted to screen for peptides exhibiting inter-group differences. In addition, all the differentially expressed peptides were examined and verified with liquid chromatography tandem mass spectrometry (LC-MS/MS).
Result An average of 77 peptide mass peaks were detected among three groups, the peaks intensities differed significantly for 10 peptides between PD patients and PD patients with COPD. Among them, eight peptides (1193.5, 1836.2, 1735.1, 1321.3, 1356.8, 2086.8, 1863.6, and 2230.9) showed increased expression and two peptides (1067.3 and 1124.4) showed decreased expression in the PD plus COPD group, in comparison with the PD group. Among the 10 differential peptides, 1193.5 and 1356.8 were identified as histidine-rich protein-1, submaxillary gland androgen-regulated protein 3B, and salivary acidic proline-rich protein 1/2.
Conclusion With WCX-MB and MALDI-TOF-MS, we have identified, from the saliva of patients with concomitant PD and COPD, differentially expressed salivary peptides that were associated with diseases. The differentially expressed peptides thus screened out show promises for being used as auxiliary biomarkers for early diagnosis of COPD.

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