QUANTITATIVE ANALYSIS OF AGE SPECIFIC VARIATION IN THE ABUNDANCE OF HUMAN FEMALE

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 人类唾液是一种富含蛋白质的、容易获得的潜在局部和系统生物标志物来源，可以监测在病理条件下发生的变化；然而，人们对与正常衰老相关的丰度变化知之甚少。在这项研究中，我们对从健康女性受试者腮腺收集的混合唾液进行了全面的蛋白质组学分析，将其分为两个年龄组(分别为20-30岁和55-65岁)。在使用多维蛋白质识别技术(MudPIT)表征腮腺唾液之前，使用疏水电荷相互作用层析分离高丰度和低丰度蛋白质。在这两个年龄组中，总共鉴定了532种蛋白质。在这些蛋白质中，有266种只在一个年龄组中被鉴定出来，而266种蛋白质在两个年龄组中都是共同的。在这两个年龄组中发现的大部分蛋白质属于防御和免疫反应类别。值得注意的是，通过G检验，组2中几种与防御相关的蛋白(如溶菌酶、乳铁蛋白和组蛋白-1)显著丰富。选择具有代表性的质谱学结果通过蛋白质印迹分析进行验证。我们的研究首次对收集自年轻和老年女性受试者的导管唾液中的差异调节蛋白进行了定量分析。这项研究支持使用高通量蛋白质组学作为一种强大的发现工具。这些结果为未来的研究提供了基础，以确定特定的唾液蛋白，这些蛋白可能与女性特有的年龄相关疾病有关。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Human saliva is a protein-rich, easily accessible source of potential local and systemic biomarkers to monitor changes that occur under pathological conditions; however, little is known about the changes in abundance associated with normal aging. In this study, we performed a comprehensive proteomic profiling of pooled saliva collected from the parotid glands of healthy female subjects, divided into two age groups 1 and 2 (20-30 and 55-65 years old, respectively). Hydrophobic charge interaction chromatography was used to separate high- from low-abundance proteins prior to characterization of the parotid saliva using multidimensional protein identification technology (MudPIT). Collectively, 532 proteins were identified in the two age groups. Of these proteins, 266 were identified exclusively in one age group, while 266 proteins were common to both groups. The majority of the proteins identified in the two age groups belonged to the defense and immune response category. Of note, several defense related proteins (e.g., lysozyme, lactoferrin and histatin-1) were significantly more abundant in group 2 as determined by G-test. Selected representative mass spectrometric findings were validated by Western blot analysis. Our study reports the first quantitative analysis of differentially regulated proteins in ductal saliva collected from young and older female subjects. This study supports the use of high-throughput proteomics as a robust discovery tool. Such results provide a foundation for future studies to identify specific salivary proteins which may be linked to age-related diseases specific to women.