臺灣博碩士論文加值系統

甲醛與反式-2,4-癸二烯醛(t,t-2,4-DDE)皆為烹飪油煙中存在的物質，家中有定時烹飪以及以烹飪為主的職業皆容易暴露在此兩種物質的環境中，故有必要分析此兩種物質的毒性作用並對此做生物性反應評估。抗氧化物的攝取對於生活節奏越加緊湊與繁忙的都會人來說是一簡便能增進建康的方法，而阿魏酸已在許多研究中被證實具有抗發炎與抗自由基等功效，為具有許多優點的抗氧化物。本研究的目的為研究甲醛與t,t-2,4-DDE對人類肺部細胞的生物效應，並藉由添加阿魏酸來評估阿魏酸的抗氧化能力。本研究利用人類肺腺癌細胞株暴露甲醛與t,t-2,4-DDE，在暴露後加入阿魏酸來培養細胞，以3- (4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay與Cell counting kit - 8 (CCK-8) assay分析細胞存活率。以MTT assay的結果顯示當細胞暴露於5至50 mM濃度的甲醛持續1至4小時細胞存活率皆顯著下降，暴露50 mM濃度的甲醛持續1小時細胞存活率減少至53%，而暴露時間增加至4小時細胞存活率更減少至13%。暴露於100至400 µM的t,t-2,4-DDE達1小時細胞存活率顯著下降，於400 µM的t,t-2,4-DDE細胞存活率減少至22%，暴露時間增長到2至4小時，暴露50 µM的t,t-2,4-DDE即造成細胞存活率顯著下降，暴露於400 µM的t,t-2,4-DDE持續2小時與4小時細胞存活率分別為24%與15%。CCK-8 assay的結果顯示當暴露於濃度10至50 mM的甲醛持續1至4小時其細胞存活率有顯著地下降，暴露於50 mM的甲醛持續1小時細胞存活率減少至69%，暴露同濃度甲醛4小時細胞存活率減少至57%。暴露於100至400 µM的t,t-2,4-DDE持續1至4小時細胞存活率會顯著下降，暴露於400 µM的t,t-2,4-DDE持續1小時細胞存活率減少至35%，暴露同濃度的t,t-2,4-DDE持續4小時細胞存活率減少至23%。阿魏酸抗氧化能力實驗結果顯示，暴露甲醛後有添加阿魏酸在細胞存活率有顯著地上升，暴露1小時30 mM的甲醛細胞存活率上升23%，其餘濃度上升約10%，而暴露於t,t-2,4-DDE後添加阿魏酸測出的細胞存活率上升幅度較小，僅在暴露1小時200 µM的t,t-2,4-DDE上升12%，其餘濃度上升皆不及10%。而隨著暴露毒物時間越長，阿魏酸對暴露在較高濃度細胞的細胞存活率影響越小，暴露2小時30至50 mM的甲醛以及暴露2小時400 µM的反式-2,4-癸二烯醛細胞存活率變化均不及5%。依本研究結果推斷，阿魏酸的抗氧化力具一定的範圍，當暴露毒物的濃度越高、暴露時間越長，以致細胞存活率過低時阿魏酸的抗氧化能力就不顯著。

Formaldehyde and trans, trans-2,4-decadienal (t,t-2,4-DDE) are two common chemicals in cooking oil fumes. Besides, formaldehyde is considered as an important indicator of indoor air quality. IARC classified formaldehyde as a group 1 carcinogen in 2012. This information reveals the importance of these two aldehydes in our daily life and their toxic effect on human being. Antioxidants play an important role preventing from diseases and improving human health nowadays. Ferulic acid is one of popular antioxidants having the abilities of anti-inflammation, scavenger of free radicals, and even anti-cancer. The aims of this study were to investigate cell viabilities of A549 human lung epithelium cells exposed to formaldehyde and trans, trans-2,4-decadienal using MTT assay and CCK-8 assay and to investigate the antioxidation effect of ferulic acid on the exposed A549 cells. In MTT assay, our results showed A549 cells exposed to formaldehyde concentrations greater than 5 mM for 1 to 4 h reduced cell viabilities. The cell viabilities reduced to 53% and 13%, when A549 cells exposed to 50 mM formaldehyde for 1 h and 4 h, respectively. Exposed to t,t-2,4-DDE concentrations ≥ 100 µM for 1h and those concentrations ≥ 50 µM for 2 to 4h also reduce cell viabilities. The cell viabilities reduced to 22% and 15%, when cells exposed to 400 µM t,t-2,4-DDE for 1h and 400 µM t,t-2,4-DDE for 4h, respectively. The same phenomena were shown in the results of CCK-8 assay. The results of ferulic acid antioxidation capability test showed ferulic acid treatment may increase cell viabilities of A549 cells. Exposed to 30 mM formaldehyde for 1h, cell viability increased 23%. As for t,t-2,4 DDE exposure, ferulic acid treatment slightly improved cell viabilities of A549 cells, in which the cell viability increased 12% for cell exposed to 200 µM t,t-2,4-DDE for 1h. The antioxidation effect of ferulic acid may not obvious while cells exposed to either high concentrations or long exposure time of formaldehyde and t,t-2,4-DDE.

致謝 I
目錄 III
表目錄 VI
圖目錄 VI
摘要 VIII
Abstract X
第一章 前言 1
1-1 研究背景 1
1-2 研究目的 3
第二章 文獻探討 4
2-1 甲醛(Formaldehyde) 4
表 2-1甲醛之化學編號與別名及物理化學性質 6
2-2 反式-2,4-癸二烯醛(Trans, trans-2,4-decadienal) 7
表 2-2反式-2,4-癸二烯醛之化學編號與別名及物理化學性質 8
2-3 阿魏酸(Ferulic acid) 9
表 2-3阿魏酸之化學編號與別名及物理化學性質 11
第三章 材料與方法 12
3-1 研究架構 12
3-2實驗器材與藥品配製 13
3-3藥品配製 14
3-4細胞之培養與處理 16
3-4-1細胞介紹與培養方式 17
3-4-2細胞培養液的配製 18
3-4-3細胞之暴露處理 19
3-5暴露危害機制與生物效應指標探討 22
3-5-1 MTT assay 23
3-5-2 CCK-8 assay 25
3-6數據分析方法 26
第四章 結果與討論 27
4-1 A549細胞暴露於甲醛之生物效應 27
4-1-1以MTT assay評估A549細胞暴露甲醛之細胞存活率 27
4-1-2以CCK-8 assay評估A549細胞暴露甲醛之細胞存活率 30
4-2 A549細胞暴露於反式-2,4-癸二烯醛之生物效應 33
4-2-1以MTT assay評估A549細胞暴露tt-DDE之細胞存活率 33
4-2-2以CCK-8 assay評估A549細胞暴露tt-DDE之細胞存活率 36
4-3 阿魏酸於A549細胞暴露醛類物質後之抗氧化能力 39
4-3-1阿魏酸對A549細胞暴露甲醛後之抗氧化能力 41
4-3-2阿魏酸對A549細胞暴露tt-DDE後之抗氧化能力 45
4-3-3阿魏酸對A549細胞暴露兩種醛類物質抗氧化能力之差異 49
4-4 研究限制 50
第五章 結論與建議 51
5-1 結論 51
5-2 建議 52
參考文獻 53

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