2012-01-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/658227摘要：由於直接與空氣接觸，呼吸道成為多環芳香烃等環境毒物的主要標的。具有生物活性的萘是最常見的多環芳香烃，廣泛存在於環境中，也因此人們時常暴露到萘。先前的研究證實，萘會造成物種和部位專一性的急性傷害；小鼠以腹腔注射暴露萘後，末端支氣管的表皮細胞會產生壞死現象；而當小鼠重複暴露萘後，標的細胞產生耐受性而不被傷害。從小鼠暴露萘而誘導細胞傷害的實驗中，我們發現活性代謝物的產生以及 glutathione 的消耗和毒性產生有關連性。為了了解萘的致毒機制及開發生物標記，廣泛檢查小分子代謝物變動是必需的研究。我們假設萘會干擾細胞內的平衡同時造成代謝物的擾亂，而觀察體液內的代謝物變動可反應細胞傷害程度；我們更可將非侵入性樣本的代謝分析應用到暴露族群，以評估環境毒物暴露所造成的健康效應。 這個研究利用易感受族群 -- 小鼠進行萘暴露引發細胞傷害機制的探討，利用核磁共振與質譜儀偵測體液代謝物後，以多變項分析來描述代謝物 profilings 的特性。萘的代謝產物及內生性代謝物都會被分析並與細胞傷害的程度做連結；小鼠暴露萘後，我們會分別使用核磁共振和質譜儀來檢測體液內親水性與親脂性小分子物質的變化，不同萘劑量及時間軸對血清及尿液中的代謝效應也會被分析，最後建議的重要分子則會以耐受動物模型進行後續的確認。本年度的研究目標著重於完成血清及尿液樣本的核磁共振資訊分析，及建立質譜儀技術分析血清脂質的平台。代謝體研究是一個強而有力的研究工具，它能提供更直接且完整的生物效應過程；這個研究希望在非侵入性樣本中找出萘暴露的生物標記，並實際應用至人類族群，這個研究也希望能發展台灣的代謝體分析平台，以期在未來利用此方法進行更多毒物、藥物及疾病的研究。 <br> Abstract: Due to direct contact with the air, the respiratory system is a primary target of environmental toxicants, such as polyaromatic hydrocarbons. Bioactivated naphthalene, the most common polyaromatic hydrocarbons, exists widely in the environment with significant human exposure. Previous studies have demonstrated that naphthalene causes species and site selective acute injury. After parenteral administration of naphthalene, bronchiolar epithelial necrosis occurs in mice. Repeated exposures to naphthalene results in target cells refractory to further injury. Formation of reactive metabolites and depletion of glutathione have been correlated with naphthalene-induced cell injury in mice. A comprehensive study examining the turbulences of small molecular metabolites is needed to understand the underlying mechanisms of naphthalene toxicity and develop biomarkers. We hypothesize that naphthalene disrupts intracellular homeostasis and causes metabolic turbulences which can be monitored and associated with cell injury in biofluids. Metabolomic measurements from non-invasive samples can be applied in exposed human populations to assess the health effects of exposure to environmental toxicants. In this proposed study, nuclear magnetic resonance (NMR)- and mass spectrometry (MS)- followed by multivariate analysis will be applied to characterize metabolic effects of naphthalene in leading to cell injury in a susceptible species, mouse. Both naphthalene metabolites and endogenous metabolites will be examined and correlated to the progress of cell injury. NMR and MS techniques will be applied to examine profiling of hydrophilic and hydrophobic metabolites, respectively, in biofluids from animals exposed to naphthalene. Time course and dose response of naphthalene-induced metabolic effects in serum and urine will be analyzed. The suggested critical molecules will be further confirmed using a tolerant animal model. In this year, we will focus on completing NMR-based metabolic analysis on urine and serum samples. Further, the platform of MS-based lipidomic analysis will be established for later application to serum samples. Metabolic techniques provide a powerful tool to describe biological processes in a more complete manner. This study will suggest the best naphthalene biomarkers in noninvasive samples with potential to be applied in the human populations. This study also intends to develop metabolic platforms in Taiwan and apply the techniques to study other toxicants, drugs, and diseases in the future.代謝體學？核磁共振質譜生物標記生物液metabolomicsnaphthalenenuclear magnetic resonancemass spectrometrybiomarkerbiofluids