Interleukin-6 and Interleukin-8 Induced by Benzo[a]pyrene、Phenanthrene、Chrysene in Human Bronchial Epithelial BEAS-2B Cell

本研究選擇具代表性的多環芳香烴化合物benzo[a]pyrene、phenanthrene、chrysene觀察人類支氣管上皮細胞（BEAS-2B cell），暴露該等多環芳香烴化合物後釋放激素interleukin-6（IL-6）及interleukin-8（IL-8）的效應。PAH暴露濃度設定為低（4.22~5.54 nM）、中（98.5~138.5 nM）、高（492.5~692.5 nM）三種範圍，暴露時間為8、12、16、24、48小時，IL-6及IL-8分析使用ELISA技術。結果發現，PAH的物種、濃度與暴露時間為影響細胞分泌IL-6與IL-8的重要變項。經暴露濃度歸一化調整後的IL-6、IL-8濃度，其單位時間的變化率隨PAH之物種而異，以benzo[a]pyrene較大。劑量-效應關係似乎呈現兩個階段，且因PAH物種而有差異：暴露benzo[a]pyrene，低劑量時IL-6、IL-8效應呈線性下降，相對較高劑量時反應隨劑量的平方而增加；而暴露phenanthrene或chrysene時，IL-6、IL-8的效應先呈線性的上升而後隨劑量的平方緩降。認為PAH造成細胞分泌細胞激素，產生發炎反應的影響可能存在，在濃度與暴露時間條件接近的情況下，不排除物種是關鍵因素。This study aimed at evaluating the induction of interleukin-6（IL-6）and interleukin-8（IL-8）while the human bronchial epithelial cells, BEAS-2B cell, exposed to authentic benzo[a]pyrene, phenanthrene, and chrysene with concentration at low（4.22~5.54 nM）, medium（98.5~138.5 nM）, and high（492.5~692.5 nM）levels. The exposure lasted for 8, 12, 16, 24, and 48 hours, before the ELISA test were applied for detecting IL-6 and IL-8. The results demonstrated that the induction of IL-6 and IL-8 varied with species of PAH, concentration and exposure time. The change rate of cytokine level per hour exposure（pg/mL-nM/hr）varied with chemical species as the levels of IL-6 and IL-8 were normalized to exposure concentration. The change rate for benzo[a]pyrene was upmost. The does-effect relationship was interpreted by using linear-quadratic model assuming that the level of IL-6 or IL-8 is related to two phases of action. For benzo[a]pyrene, the level of IL-6 and IL-8 were negatively proportional to dose at the lower dose exposure and the level of IL-6 and IL-8 were positively proportional to the square of dose at the higher dose exposure. For exposure to phenanthrene or chrysene, the levels of IL-6 and IL-8 positively increase with the increment of dose at lower dose exposure and then slightly decrease with the square of dose at higher dose exposure. The induction of proinflammatory cytokines in epithelial cell likely occurs due to exposure to polynuclear aromatic hydrocarbons, and the effects mainly depend on chemical species as the other exposure modifiers such as exposure concentration and exposure time are identical.致謝I要IIbstractIII錄IV列VI列VII一章 緒論1. 前言1. 研究目的2. 研究架構3二章 文獻回顧4. 環境中的多環芳香烴化合物4. 空氣污染對氣喘發生之影響4. 呼吸系統的發炎反應5. 多環芳香烴化合物對健康之影響6. 人類呼吸系統免疫機制8三章 材料與方法10. 材料10. 方法12學物質多環芳香烴的濃度配製12胞暴露實驗12胞繼代培養12胞密度計數15種（Seeding）16胞暴露17LISA分析17. 資料處理19四章 結果與討論20. 暴露時間、暴露濃度對BEAS-2B cell分泌IL-6與IL-8的影響20. 細胞分泌IL-6、IL-8對於暴露PAH單位濃度、暴露時間與PAH物種的反應21. IL-6、IL-8與PAHs暴露劑量（時間×濃度）的關係24. PAH大氣樣品對細胞分泌IL-6、IL-8濃度影響25. 研究的限制與建議26考文獻28錄列一.多環芳香烴的濃度配製34二.BEAS-2B細胞株詳細資料34三.BEAS-2B cell暴露benzo[a]pyrene分泌IL-6、IL-8之結果35四.BEAS-2B cell暴露Phenanthrene分泌IL-6、IL-8之結果37五.BEAS-2B cell暴露Chrysene分泌IL-6、IL-8之結果39六.BEAS-2B cell分泌IL-6與PAH物種、濃度分組及暴露時間的關係41七.BEAS-2B cell分泌IL-8與PAH物種、濃度分組及暴露時間的關係42八.IL-6濃度與暴露時間的指數關係43九.IL-8濃度與暴露時間的指數關係44列一.BEAS-2B cell 分泌IL-6與PAH物種、濃度分組及暴露時間的關係45二.BEAS-2B cell 分泌IL-8與PAH物種、濃度分組及暴露時間的關係46三.高濃度PAH暴露下IL-6反應與暴露時間的關係47四.中濃度PAH暴露下IL-6反應與暴露時間的關係48五.低濃度PAH暴露下IL-6反應與暴露時間的關係49六.高濃度PAH暴露下IL-8變化與暴露時間的關係50七.中濃度PAH暴露下IL-8濃度變化與暴露時間的關係51八.低濃度PAH暴露下IL-8濃度變化與暴露時間的關係52九.IL-6效應與PAH暴露劑量(濃度×時間)的關係53十.IL-8效應與PAH暴露劑量(濃度×時間)的關係5