Rudney J D, Staikov R K, Johnson J D
Department of Diagnostic and Biological Sciences, School of Dentistry, 17-252 Moos Tower, 515 Delaware St. SE, University of Minnesota, Minneapolis, MN 55455, USA.
Arch Oral Biol. 2009 Jan;54(1):91-100. doi: 10.1016/j.archoralbio.2008.08.007. Epub 2008 Sep 18.
In previous studies, we defined groups of subjects with opposite salivary function. Group membership was associated with clinically relevant outcomes. High aggregation-adherence (HAA) groups showed lower levels of caries, supragingival plaque, total streptococci, and Tannerella forsythensis than low high aggregation-adherence (LAA) groups. In this study, we used a proteomic approach to search for biomarkers which could be useful as risk indicators for those outcomes.
Clarified resting whole saliva from each of 41 HAA and LAA subjects was separated by preparative isoelectric focusing. Fractions showing the most distinctive protein profiles were pooled into four sets (pI 3-3.5, pI 4-4.7, pI 5.7-7.7, pI 10-11.5). Each pool then was compared by SDS-PAGE. Image analysis software was used to quantify matched bands. Partial least squares analysis (PLS) was used to determine which of the 65 bands from all four pools were the best predictors of group membership, caries, total plaque, total streptococci, and T. forsythensis counts. Those bands were identified by mass spectroscopy (MS-MS).
Two bands consistently were strong predictors in separate PLS analyses of each outcome variable. In follow-up univariate analyses, those bands showed the strongest significant differences between the HAA and LAA groups. They also showed significant inverse correlations with caries and all the microbiological variables. MSMS identified those bands as statherin, and a truncated cystatin S missing the first eight N-terminal amino acids.
Levels of statherin and truncated cystatin S may be potential risk indicators for the development of caries and other oral diseases.
在先前的研究中,我们定义了唾液功能相反的受试者组。组的划分与临床相关结果相关。高聚集黏附(HAA)组的龋齿、龈上菌斑、总链球菌和具核梭杆菌水平低于低高聚集黏附(LAA)组。在本研究中,我们采用蛋白质组学方法寻找可作为这些结果风险指标的生物标志物。
对41名HAA和LAA受试者的静息全唾液进行澄清,通过制备性等电聚焦进行分离。显示出最独特蛋白质谱的馏分被合并为四组(pH值3 - 3.5、pH值4 - 4.7、pH值5.7 - 7.7、pH值10 - 11.5)。然后通过SDS - PAGE对每组进行比较。使用图像分析软件对匹配条带进行定量。采用偏最小二乘法分析(PLS)来确定来自所有四组的65条带中哪些是组划分、龋齿、总菌斑、总链球菌和具核梭杆菌计数的最佳预测指标。通过质谱(MS - MS)鉴定这些条带。
在对每个结果变量的单独PLS分析中,两条带始终是强预测指标。在后续的单变量分析中,这些条带在HAA组和LAA组之间显示出最强的显著差异。它们还与龋齿和所有微生物学变量呈显著负相关。MS - MS将这些条带鉴定为富含组氨酸的糖蛋白和缺失前八个N端氨基酸的截短胱抑素S。
富含组氨酸的糖蛋白和截短胱抑素S的水平可能是龋齿和其他口腔疾病发生的潜在风险指标。