- 學位論文
偵測呼出氣體凝結液中所含反應性含氧物種作為指標在環境衛生上的應用──可行性探討
Measurement of Reactive Oxygen Species in Exhaled Breath Condensate as an Index in Environmental Health─A Feasibility Study
摘要
分析呼出氣體凝結液(exhaled breath condensate; EBC)中的成分做為診斷工具，由於其採樣方式簡單且具有非侵入的特性，因此近年來格外受到注意。例如：部分研究針對呼出氣體凝結液中的小分子(如：H2O2、8-isoprostane、NO衍生的產物等)與氧化壓力或呼吸系統疾病的相關性進行探討。由於現有文獻多數未詳述對於EBC採樣及分析所得數據的品管，導致在各文獻的結果比較上相對困難。 本研究的目標在於改進現有的呼出氣體凝結液中反應性含氧物種(reactive oxygen species, ROS)，簡稱EBC-ROS的採樣分析方法，並探討各項可能影響分析結果的因素。進一步以本研究的採樣分析方法用於測量及比較抽菸者與非抽菸者的EBC-ROS濃度。 本研究以一個改良後的EBC採樣系統進行EBC的收集，使用DCFH2-HRP化學螢光法來偵測呼出氣體凝結液中ROS濃度。EBC的收集方面，於採樣管最前端加裝一個如衝擊瓶形式的唾液捕集裝置，此裝置可有效避免EBC受到唾液的污染。將乾式氣體流量計架設於採樣系統的末端，用於準確測量採集EBC時受試者的呼氣體積。以上的兩個裝置有效改善數據的品質，且在重複樣品試驗的結果顯示其變異程度低於10%。 除了基本的QA/QC試驗以外，其他如不同採樣體積以及同日不同時段採樣對於EBC中ROS濃度的影響，亦於本研究中探討。以呼氣總體積60公升及40公升來收集EBC，二者的ROS濃度濃度並無不同。於18:00所測得的平均EBC-ROS濃度顯著高於10:30，12:00以及15:00所測得的數值；造成此種變異情形可能和受試者一天中的活動或污染物暴露狀況有關。 抽菸者於抽菸後，呼出氣體凝結液中ROS濃度顯著比抽菸前高，但是抽菸後三十分鐘測得的濃度則與抽菸前沒有差異；最後偵測8名輕度抽菸者和18名非抽菸者的EBC中ROS濃度，發現兩組受試者的數值相近，僅有一名有嚼檳榔習慣之抽菸者得到較高的測值。此差異點可在未來的研究中作進一步的探討。
並列摘要
Analysis of exhaled breath condensate (EBC) components for diagnostic purpose has received attention in recent years because the sampling method is very simple and noninvasive. For example, several studies measured small molecules in EBC (H2O2, 8-isoprostane, and NO-derived products, etc.) to correlate with oxidative stress or disease in pulmonary system. None of the EBC sampling and analysis methods in current literatures has detailed description about how they control their data measurement quality however, thus makes the comparison of their results relatively difficult. This study aimed at improving the EBC-reactive oxygen species (EBC-ROS) collection and analysis method, and investigating factors that might influence the quality of the data measurement. The method then was used to measure and compare the EBC-ROS concentration in nonsmokers and smokers. An improved EBC collecting system was used to collect EBC, and the ROS concentration in EBC was measured using a DCFH2-HRP chemical fluorescent method. For EBC collection, a simple impinger type saliva trapping device was connected to the front end of the EBC collecting tubing, this device can effectively prevent the pollution of EBC by saliva. To measure exhaled air volume of each EBC sample collected accurately, a dry test gas meter was added at the end of the EBC collecting device. Both these modification greatly improved data quality, and the duplicate sample analysis indicated less than 10% variation can be obtained. In addition to the basic QA/QC tests, factors including sampling volume and the sampling time within a day were also examined for their effect on the ROS concentration measurement. No difference was observed in ROS concentration in EBC collected either using 60 or 40 liters of exhaled air. The mean EBC-ROS concentration was significantly higher at 18:00 than at 10:30, 12:00, and 15:00. This within-day variation at late afternoon may be either due to the test subjects’ one-day activities or due to the whole day’s exposure of pollutants. Smokers immediately after smoking showed a significant increase in EBC-ROS concentration as comparing with before smoking, but no difference after half an hour of stop smoking. EBC collected from eight current light smokers and eighteen non-smokers have similar ROS concentration levels, except one who has the habit of betel nut chewing, this should worth further study.
參考文獻
延伸閱讀
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