Core--Eicosanoid Analysis

产品说明：（申请人的描述）类花生酸分析核心的具体目标是为该计划项目资助的研究人员进行复杂生物液体和组织中的类花生酸分析。本计划项目补助金中提出的四个项目涉及研究环氧合酶和类花生酸在结直肠肿瘤中的作用，并将需要定量测定前列腺素（PGs）。将通过采用毛细管气相色谱（GC）/电子捕获负离子化学电离（NICI）质谱（MS）的方法，使用由核心负责人开发且目前正在使用的稳定同位素稀释技术，对细胞孵育物以及动物和人体组织中的类二十烷酸进行定量。 参与该计划项目资助的研究人员使用这些测定已经允许进行一些基本观察，这些观察有助于描述类花生酸在与胃肠道恶性肿瘤相关的病理生理过程中的作用。此外，核心还将开发GC/NICI MS测定法，以测量由环加氧酶产生的双环内过氧化物中间体PGH 2衍生的化合物12-羟基-十七碳三烯酸（12-HHT）。 该试验将用于项目4（Dr. Marnett），以进一步探索12-HHT在环氧合酶过度表达情况下的作用。因此，类二十烷酸核心的总体目标是为类二十烷酸的定量分析提供技术支持和实施高灵敏度方法。

DESCRIPTION: (Applicant's Description) The specific goals of the Eicosanoid Analysis Core are to carry out the analyses of eicosanoids in complex biological fluids and tissues for investigators in this Program Project Grant. The four projects proposed in this Program Project Grant involve research examining the role of the cyclooxygenase and eicosanoids in colorectal neoplasia and will require quantification of prostaglandins (PGs). Eicosanoids will be quantified in cellular incubations and animal and human tissues by methodology employing capillary gas chromatography (GC)/electron capture negative ion chemical ionization (NICI) mass spectrometry (MS) using stable isotope dilution techniques that were developed by the Core Leader and which are currently in use. Use of these assays by investigators involved in this Program Project Grant has already allowed a number of fundamental observations to be made that have helped to delineate the role of eicosanoids in pathophysiological processes related to gastrointestinal malignancy. In addition, the Core will also develop a GC/NICI MS assay to measure the compound 12- hydroxy-heptadecatrienoic acid (12-HHT) derived from the cyclooxygenase-generated bicycloendoperoxide intermediate PGH2. This assay will be utilized in Project 4 (Dr. Marnett) to further explore the role of 12-HHT in situations of cyclooxygenase overexpression. The overall objectives of the Eicosanoid Core, therefore, are to provide technical support and implementation of highly sensitive methodology for the quantitative analysis of eicosanoids.