研究微粒帶電性質與呼吸毒性之關聯: 以小鼠暴露奈米黑碳微粒實驗為例

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沈妤芳(Yu-Fang Shen) 查詢紙本館藏

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環境工程研究所

論文名稱

研究微粒帶電性質與呼吸毒性之關聯: 以小鼠暴露奈米黑碳微粒實驗為例
(Inhalation Exposure Study of Soot Nanoparticles with Positive, Neutral and Negative Charges in Mice)

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摘要(中)

黑碳是交通汙染源的指標之一，由於其不規則的構形易夾帶有機物、無機物甚至是有毒物質，若人體長期吸入會增加致癌率。近年來，許多研究探討暴露在奈米黑碳微粒環境下對於健康的影響，但針對帶電微粒所導致的健康傷害目前尚未有清楚的了解，因此本研究針對帶電奈米黑碳微粒對於呼吸道健康進行研究，並建立一套穩定的奈米黑碳微粒暴露系統。實驗將BALB/c雌性小鼠暴露於奈米黑碳微粒且分為帶正電與帶負電兩組，每組各有四種條件，分別為高濃度、低濃度、電性中和及HEPA控制組，每日暴露八小時且持續一周。利用電移動度分析儀 (DMA) 篩選微粒粒徑為70 nm，後端以電移動度掃描分徑器 (SMPS) 確認篩選粒徑、奈米微粒表面積偵測器 (NSAM) 量測微粒沉積在肺部的表面積、氣膠電流計 (AE) 確認微粒的電性、黑碳監測儀 (AE31)量測黑碳濃度與同時收集濾紙進行成份與構形分析。綜合以上儀器可隨時監測系統的穩定性，並且相互比較量測值的準確性。暴露後一週犧牲並收集小鼠的肺部灌洗液、血液與肺部組織，評估發炎反應與肺部組織的破壞程度 (BALF Total Protein、BALF LDH、Serum IL-6、Lung 8-Isoprostane、Lung Caspase-3、Lung BPDE)。BALF-LDH、Lung 8-Isoprostane、 Serum IL-6皆與黑碳的質量濃度、肺部沉積表面積濃度與電流量有良好的線性關係。尤其BALF-LDH的分析具有最佳的相關性，並可觀察出在低暴露劑量的範圍下，比起正電與電性中和黑碳奈米微粒，負電黑碳奈米微粒所造成的生物毒性較低。

摘要(英)

Soot particle is one of the indicators of traffic pollution. It can contain organic, inorganic or even toxic substances easier due to its irregular structure. Exposure to soot particle significantly increased the risk of lung cancer. In recent years, many studies investigate the health effects after exposure to soot particles, but for the charged particles is not clear understanding yet. Therefore, this study establish a stable soot particle exposure system and study the biological responses of mouse after exposing to the charged soot particles. BALB/c female mice were exposed to soot particles into positively and negatively charged groups, each have four conditions (high concentration, low concentration, electrical and control group), and exposed for eight hours a day and last for a week. Soot particle size were select 70 nm by Differential Mobility Analyze (DMA) and confirm particle size by Scanning Mobility Particle Sizer (SMPS), Nanoparticle Surface Area Monitor (NSAM) measure the surface area of the particles deposited in the lungs, Aerosol Electrometer (AE) confirm the electric charge of particles, Aethalometer (AE31) and Optical Transmissometer (OT21) measure the concentration of soot particle. At the same time, filter were collect soot particle for composition and structure analysis. Based on the above instruments, they can monitor the stability of the system and the compare the accuracy of the measurement data. After exposure, we collected lung tissue, serum and bronchoalveolar lavage fluid of mouse to study the biological responses (BALF-Total Protein、BALF-LDH、Serum-IL-6、Lung-8-Isoprostane、Lung-Caspase-3、Lung-BPDE). The levels of BALF-LDH, Lung 8-Isoprostane and Serum IL-6 are well linearly correlated to BC mass concentration, lung deposition surface area concentration and charge current. Especially for BALF-LDH, the correlations are almost perfect linear. Furthermore, the responses for negative charged soot particles are significantly lower than both neutral and positive charge particles (especially in low exposure dosage range).

關鍵字(中)

★ 帶電微粒
★ 呼吸暴露
★ 奈米黑碳微粒

關鍵字(英)

★ Charged particles
★ Inhalation exposure
★ Soot Nanoparticles

論文目次

摘要 I
Abstract II
目錄 IV
圖目錄 VII
表目錄 IX
第一章前言 1
1-1研究動機 1
1-2研究目的 2
第二章文獻回顧 3
2-1奈米微粒的生物毒性 3
2-1-1 微粒粒徑 3
2-1-2 微粒構形 5
2-1-3 微粒電性 6
2-2奈米微粒的健康效應 8
2-3黑碳的特性 10
2-3-1 黑碳的來源 10
2-3-2 黑碳的形成 13
2-3-3 黑碳的毒性 17
2-4黑碳的健康效應 21
2-4-1 交通汙染源 21
2-4-2 職業傷害 22
2-4-3 實驗室暴露 23
第三章研究方法 24
3-1研究架構 24
3-2實驗流程 24
3-3研究設計 25
3-4暴露實驗 26
3-3-1 產生源系統 27
3-3-2 篩分系統 34
3-3-3 暴露系統 34
3-3-4 量測系統 37
3-4毒理分析 39
3-4-1以Bradford 法進行肺部灌洗液蛋白質定量 40
3-4-2肺部灌洗液與乳酸脫氫酶LDH評估 41
3-4-3血液與細胞激素IL-6 41
3-4-4以Bradford 法進行肺部組織蛋白質定量 43
3-4-5肺部組織與氧化壓力 44
3-4-6肺部組織與Caspase-3 45
3-4-7肺部組織與BPDE 46
3-4統計分析 47
3-5濾紙分析 48
第四章結果與討論 51
4-1量測結果 51
4-1-1 系統的穩定性與燃燒狀態 51
4-1-2 微粒沉積在肺部的表面積濃度 57
4-1-3 微粒構形與電性 59
4-1-4 微粒成分與毒性分析 63
4-2毒理分析 70
4-2-1 肺部灌洗液毒理分析 (Total protein、LDH) 70
4-2-2 血液毒理分析 (IL-6) 72
4-2-3 肺部組織毒理分析(8-Isoprostane、Caspase-3、BPDE) 72
4-2-4 小鼠體重分析 75
4-2-5 小鼠病理切片分析 76
4-3結果討論 78
4-3-1 微粒沉積於肺部的表面積對於呼吸毒理的關聯性 78
4-3-2 微粒電性對於呼吸毒理的關聯性 80
4-3-3 微粒成分對於呼吸毒理的關聯性 83
4-3-4 多元迴歸 87
第五章結論 90
5-1 系統狀態 90
5-2微粒構形及成分 90
5-3毒理分析 91
5-4量測結果與毒理分析之關聯性 91
參考文獻 92
附錄 97
口試委員意見回覆 100

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指導教授

蕭大智(Ta-Chih Hsiao)

審核日期

2015-12-2

推文