臺灣博碩士論文加值系統

此研究目的為探究噴漆作業中所產生的噴漆懸浮微粒是否會經由呼吸道進入人體並沉積在肺部進而造成健康上的危害及其與慢性阻塞型肺病COPD之相關性，此實驗將所採樣之噴漆懸浮微粒依其顆粒粒徑分為PM10-3.2、PM3.2-1以及PM1三種大小，並計算可能沉積於肺部之濃度後與人類肺支氣管上皮細胞BEAS-2B進行了細胞毒性以及細胞屏障完整度的測試。在細胞存活率測試MTT中，發現到三種大小的PM皆會對細胞造成損害，其中PM10-3.2於高劑量即100 μg/cm2造成了25%的細胞死亡並且與正常組相比具有一顆星的顯著差異，而在PM3.2-1以及PM1兩種顆粒大小則於中高劑量即50 μg/cm2與100 μg/cm2皆造成10-20%的細胞死亡且與正常組相比具有兩顆以及三顆星的顯著差異，說明顆粒粒徑越小所造成的細胞死亡就越多。其後對於細胞進行發炎因子IL-6以及IL-8蛋白表現量的測定，也發現這些顆粒會使細胞產生發炎反應，其中PM10-3.2之顆粒於高劑量下會使得細胞大量分泌IL-6，其濃度為700 pg/ml，而正常組為200 pg/ml，PM3.2-1則於高劑量下使IL-6提高至800 pg/ml，而PM1則在低劑量以及高劑量皆使得IL-6提高至900 pg/ml以上，而在IL-8中亦是如此，因此初步判定懸浮微粒可能透過造成細胞發炎而導致細胞死亡且PM1相較於其他兩種大小之顆粒更容易對細胞產生影響。其後針對COPD的相關標的蛋白如緊密連結蛋白ZO-1、AAT進行表達測定，也針對皮膜屏障完整度進行的跨膜電阻的測試後發現，顆粒確實會使ZO-1、AAT的表現量降低，皮膜屏障的完整性也受到破壞，大幅提高了罹患COPD的可能性。因此，在工作場所中配戴適當的護具以降低懸浮微粒的威脅，是必須的。

The airborne particulate matters (PM) has been a serious issue for decades. Apart from environmental PM, PM may also be produced indoor during spray painting. Workers who were exposed to these PM may be in the risks of respiratory diseases like chronic obstructive pulmonary disease (COPD). In this study, the spray paint particles were separated in three groups depending on their size, PM10-3.2, PM3.2-1 and PM1. The cytotoxicity of the spray paint PM toward human bronchial epithelial cell BEAS-2B was investigated. Results showed that the smaller PM were, the more serious cytotoxicity and inflammation would be caused. Also, we found PM damaged the cell in dose dependent manner. Furthermore, the biomarkers of COPD like tight junction protein zonula occludens 1 (ZO-1), a-1-antitrypsin (AAT) and epithelial resistance (ER) were decreased as well. The evidences demonstrated that exposure to the spray paint PM may increase the risk of getting COPD. Therefore, it is necessary for the workers to wear appropriate equipment during spray painting.

摘要... i
Abstract....... ii
誌謝... iii
目錄... iv
表目錄.. vi
圖目錄.. vii
縮寫對照表...... viii
第一章 緒論..... 1
第二章 文獻回顧. 2
2.1 噴漆成分.... 2
2.2 懸浮微粒 (PM)....... 2
2.3 懸浮微粒 PM 之暴露途徑....... 4
2.3.1 眼部途徑.. 4
2.3.2 皮膚途徑.. 4
2.3.3 呼吸道途徑........ 5
2.4 懸浮微粒PM之毒性機制. 6
2.5 慢性阻塞型肺病COPD.. 8
第三章 材料與方法....... 9
3.1 實驗流程.... 9
3.2 顆粒採樣.... 9
3.2.1 顆粒萃取.. 9
3.3 細胞培養.... 10
3.4 細胞存活率測試 (MTT)........ 10
3.5 活性氧化物測定 (ROS)........ 10
3.6 發炎因子 IL-8 測定.......... 10
3.7 發炎因子 IL-6 測定.......... 10
3.8 緊密連接蛋白 ZO-1測定....... 11
3.9 α-1-antitrypsin (AAT) 測定......... 11
3.10 跨膜電阻 TEER 測定......... 11
第四章 結果與討論....... 12
4.1 顆粒採樣.... 12
4.2 細胞存活率 (MTT).... 17
4.3 活性氧化物 (ROS).... 18
4.4 細胞激素IL-6........ 19
4.5 細胞激素IL-8........ 20
4.6 緊密連接蛋白 ZO-1... 21
4.7 跨膜電阻TEER.....22
4.8 α-1型抗胰蛋白酶 (AAT)....... 23
第五章 結論..... 25
參考文獻 ......26
Extended Abstract...... 32

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