Blanco-Pintos Triana, Regueira-Iglesias Alba, Kuz Iryna, Sánchez-Barco Alba, Seijas-Otero Noelia, Chantada-Vázquez María Del Pilar, Balsa-Castro Carlos, Tomás Inmaculada
Oral Sciences Research Group, Special Needs Unit, Department of Surgery and Medical-Surgical Specialties, School of Medicine and Dentistry, Universidade de Santiago de Compostela, Health Research Institute of Santiago (IDIS), Santiago de Compostela, Spain.
Proteomic Unit, Health Research Institute of Santiago de Compostela (IDIS), Santiago de Compostela, Spain.
J Periodontol. 2025 Mar;96(3):217-229. doi: 10.1002/JPER.24-0062. Epub 2024 Sep 16.
Few investigations evaluated smoking's impact on the periodontal proteome. Therefore, this study aimed to analyse the influence of tobacco on the overall periodontal proteome and the differential expression of gingival crevicular fluid (GCF) proteins using sequential window acquisition of all theoretical mass spectra (SWATH-MS).
GCF samples were collected from 40 periodontitis subjects (stages III-IV). These were separated based on smoking status into smokers (17), ex-smokers (10), and non-smokers (13). Samples were analysed using SWATH-MS, and proteins were identified using the UniProt human-specific database. Data are available via ProteomeXchange with the identifier PXD043474. Principal component analysis (PCA) was employed to examine the spectral mass distribution of the proteome. Protein expression was different for a p-value <0.05 and a log2 fold change ≥0.3 (upregulated) or ≤-0.3 (downregulated).
The distribution of overall proteome did not differ between non-smokers, smokers, and ex-smokers. Considering protein expression, 23 were differentially expressed in smokers vs. non-smokers (16 upregulated and 7 downregulated), 17 in ex-smokers vs. non-smokers (2 upregulated and 15 downregulated), and only 8 in smokers vs. ex-smokers (7 upregulated and 1 downregulated). Smoking increased the expression of proteins related to epithelial hyperkeratinization (keratins type II cytoskeletal 4, type I cytoskeletal 13 and type I cytoskeletal 19, cornulin, and fatty acid-binding protein 5). However, multiple immunoglobulins were underexpressed when comparing smokers and ex-smokers to non-smokers.
Although smoking does not significantly modify the overall GCF proteome associated with periodontitis, it alters the expression of several proteins compared to never-smokers and ex-smokers.
Smoking is a critical risk factor for the development and progression of periodontitis. However, evidence of the effect of smoking on the subgingival proteome is scarce. Therefore, this study aimed to determine the impact of smoking on the overall proteome and differential expression of gingival crevicular fluid (GCF) proteins using the sequential window acquisition of all theoretical mass spectra (SWATH-MS) proteomic technique. For this purpose, GCF samples were collected from 40 subjects with periodontitis, of which 17 were smokers, 10 were ex-smokers, and 13 were non-smokers. These samples were analysed by SWATH-MS, and proteins were identified using the UniProt human-specific database. Analysis of the overall proteome showed that its distribution was not significantly different between smokers, ex-smokers, and non-smokers. However, several proteins were found to be differentially expressed according to the smoking status. Smoking can increase the expression of several keratins and proteins related to hyperkeratinization of the epithelium. However, in ex-smokers, these proteins return to similar levels to those of non-smokers. Moreover, smoking may induce a lower expression of proteins related to adaptive immunity, such as immunoglobulins. This immunosuppressive effect may persist in ex-smokers.
很少有研究评估吸烟对牙周蛋白质组的影响。因此，本研究旨在利用所有理论质谱的序列窗口采集技术（SWATH-MS）分析烟草对整体牙周蛋白质组的影响以及龈沟液（GCF）蛋白质的差异表达。
从40名牙周炎患者（III-IV期）中收集GCF样本。根据吸烟状况将这些患者分为吸烟者（17例）、戒烟者（10例）和非吸烟者（13例）。使用SWATH-MS分析样本，并使用UniProt人类特异性数据库鉴定蛋白质。数据可通过ProteomeXchange获得，标识符为PXD043474。采用主成分分析（PCA）来检查蛋白质组的质谱分布。当p值<0.05且log2倍数变化≥0.3（上调）或≤ -0.3（下调）时，蛋白质表达存在差异。
非吸烟者、吸烟者和戒烟者之间整体蛋白质组的分布没有差异。在蛋白质表达方面，吸烟者与非吸烟者相比有23种蛋白质差异表达（16种上调，7种下调），戒烟者与非吸烟者相比有17种（2种上调，15种下调），吸烟者与戒烟者相比仅有8种（7种上调，1种下调）。吸烟增加了与上皮过度角化相关的蛋白质的表达（II型细胞骨架角蛋白4、I型细胞骨架角蛋白13和I型细胞骨架角蛋白19、兜甲蛋白和脂肪酸结合蛋白5）。然而，将吸烟者和戒烟者与非吸烟者相比时，多种免疫球蛋白表达不足。
虽然吸烟不会显著改变与牙周炎相关的整体GCF蛋白质组，但与从不吸烟者和戒烟者相比，它会改变几种蛋白质的表达。
吸烟是牙周炎发生和发展的关键危险因素。然而，吸烟对龈下蛋白质组影响的证据很少。因此，本研究旨在使用所有理论质谱的序列窗口采集技术（SWATH-MS）蛋白质组学技术来确定吸烟对整体蛋白质组以及龈沟液（GCF）蛋白质差异表达的影响。为此，从40名牙周炎患者中收集GCF样本，其中17名是吸烟者，10名是戒烟者，13名是非吸烟者。这些样本通过SWATH-MS进行分析，并使用UniProt人类特异性数据库鉴定蛋白质。对整体蛋白质组的分析表明，吸烟者、戒烟者和非吸烟者之间其分布没有显著差异。然而，发现几种蛋白质根据吸烟状况存在差异表达。吸烟可增加几种角蛋白和与上皮过度角化相关的蛋白质的表达。然而，在戒烟者中，这些蛋白质的水平恢复到与非吸烟者相似的水平。此外，吸烟可能会导致与适应性免疫相关的蛋白质（如免疫球蛋白）表达降低。这种免疫抑制作用可能在戒烟者中持续存在。
