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  • 學位論文

利用核磁共振光譜儀的代謝體學來研究萘對小鼠組織的影響

1H NMR-based Metabolomics to Study Effects of Naphthalene on Mouse Tissues

指導教授 : 林靖愉

摘要


萘是常見的多環芳香烴,萘廣泛的運用在生活製品中且因為它的易揮發性,使人類於日常生活中容易不經意地暴露過量的萘,然而萘的毒性作用機制目前仍不完全清楚。代謝體學透過完整地分析內生性代謝物的變化,可以了解外來物在生物體內的生物效應,藉由核磁共振光譜儀的代謝體學方法能全面的研究萘對小鼠各組織代謝的影響並探討萘的毒性機制。 本研究利用腹腔注射ICR品種的小鼠進行劑量反應實驗,共三組:對照組(橄欖油)、100 mg/kg及200 mg/kg的萘劑量,接著利用核磁共振光譜儀分析、主成分分析(PCA)及正交偏最小二乘判别分析(OPLS-DA)來研究肺、肝及腎組織的代謝體變化。從研究結果發現,三種組織經由實驗處裡的代謝反應皆可由主成分分析加以區分,並經傳統統計檢驗發現許多重要代謝物呈現顯著變化;萘對肺組織的影響可能包括細胞膜結構的損害及粒線體能量代謝的干擾,分別反映在Glycerophosphocholine量的下降及Succinate的上升,而萘對肝及腎組織的影響與抗嗜電子性化合物(electrophiles) 反應機制有很高的相關,肝組織中Glutamine和UDP-glucose隨萘暴露劑量提高呈現顯著上升,腎組織中Glutathione的量也隨萘暴露劑量增加呈顯著上升,這可能表示萘暴露啟動了肝及腎組織的解毒機制。 我們的研究結果顯示萘對小鼠的毒性作用不只會影響主要標的器官肺,對於肝及腎組織的代謝體同樣會造成影響,透過代謝物的影響可以部分解釋萘對小鼠的病理結果,本研究也證明了核磁共振儀的代謝體學研究方法對於了解毒性作用機制是相當有力的工具。

並列摘要


Naphthalene, a primary polycyclic aromatic hydrocarbon (PAH), widely spread in the environment was hypothesized to relate with adverse health effects. In order to clarify the underlying mechanisms of naphthalene-induce toxicity, extensive studies are required. Therefore, this study intended to investigate the mechanisms of naphthalene-induce toxicity in various tissues of mice. We used high through-put 1H NMR-based metabolomics to study effects of naphthalene on mouse tissues. Dose-response experiments (0, 100 and 200 mg/kg naphthalene, ip) were carried out on male ICR mice. Both hydrophilic and hydrophobic metabolites from the lung, liver and kidney tissues were extracted and analyzed by 1H and p-J-resolved NMR followed by principal component analysis (PCA) and orthogonal projections to latent structures discriminant analysis (OPLS-DA). From our results, the metabolic effects of naphthalene on lung, liver, and kidney demonstrated dose-dependent effects. Our results suggest that naphthalene-induced lung injures were associated with cellular membrane damages and turbulence of energy metabolisms, where declination of glycerophosphocholine and increase of succinate was found in the lung. In the liver, glutamine and UDP-glucose were increased in the 200 mg/kg exposure group. Besides, detoxification nucleophile glutathione, also demonstrated to be increased depending on naphthalene dose in the kidney. Through this study, not only the metabolome of target organ lung but metabolome of liver and kidney were found to respond to naphthalene intervention. The changes of these metabolites may partially explain the pathological response of naphthalene exposure. In conclusion, 1H NMR-based metabolomics is a useful tool in understanding the mechanism of xenobiotic induced toxicities.

並列關鍵字

naphthalene mouse lung toxicity liver kidney NMR metabolomics

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