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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 鄭尊仁(Tsun-Jen Cheng) | |
| dc.contributor.author | Tzu-Hsuan Kuo | en |
| dc.contributor.author | 郭子瑄 | zh_TW |
| dc.date.accessioned | 2022-11-25T03:05:12Z | - |
| dc.date.available | 2023-01-01 | |
| dc.date.copyright | 2021-07-20 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-07-12 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/81857 | - |
| dc.description.abstract | 塑膠微粒為近年來興起的全球議題，其被定義為粒徑小於5毫米的聚合物，廣泛的分佈在環境中，已在各大海洋、湖泊及冰川檢測到塑膠微粒，因其不易降解的特性，將隨著食物鏈累積在生物體，除了海洋生物，已有研究顯示哺乳類，例如海豚、海龜及海鷗都有檢測出塑膠微粒。暴露到塑膠微粒的途徑有皮膚吸收、呼吸道吸入及食道攝入這三種途徑，其中，攝入(Ingestion)被認為是人類主要暴露塑膠微粒的途徑。 相關研究過去多著重於微米級塑膠微粒(>1 µm)，顯示塑膠微粒在攝入後將會被腸胃道吸收並且累積在肝臟、脾臟及腎臟等器官，並且將會造成氧化壓力及發炎反應，但結果並不一致，對於哺乳類毒性的研究也很有限。且已有研究顯示越小的塑膠微粒在生物體內累積後的清除效率越低，而在體外細胞實驗，在相同重量濃度下較小的塑膠微粒其因為有較大的總表面積，會產生較大的細胞毒性。為進一步瞭解次微米級塑膠微粒(<1 µm)的健康效應，本研究探討次微米塑膠微粒在小鼠腸道的吸收、在體內的分佈及毒理效應。 本研究使用粒徑800 nm與200 nm的Nile Red螢光聚苯乙烯微珠，以胃管灌食6週大C57BL/6母鼠，為比較200 nm與800 nm微粒在組織的分佈，每次灌食109顆，每週三次連續灌食四週。暴露後24小時尿液以全代謝籠於暴露期間隔週收集，周邊血液於犧牲後由心臟收集，同時採集腸道及其他器官，包括肝臟、脾臟、腎臟及肺臟。使用螢光顯微鏡及全景組織細胞分析儀(TissueFAXS)觀察塑膠微粒在組織的分佈及累積，及使用流式細胞儀(Flow cytometry)來檢測血液、尿液及將器官組織消化過後的組織消化液，以獲得單位微粒數目濃度。 另外檢測生物指標，將部份尿液做四種全身性氧化壓力(Oxidative stress)的濃度檢測，包括8-OHdG、8-NO2Gua、8-IsoPF2a及HNE-MA，及將肺臟及腎臟組織檢測四種發炎因子(Cytokine)的含量，包括Interferon- γ (IFNγ)、Interleukin- 1β (IL-1β)、Interleukin-6 (IL-6)與Tumor Necrosis Factor- α (TNFα)。 在以胃管灌食小鼠四週後，次微米塑膠微粒會穿越腸胃道屏障進入血液循環，並進一步累積在肝臟、脾臟及腎臟，平均每個器官將會有102 ~ 103的微粒累積，並且肺臟在兩者次微米塑膠微粒皆是最主要微粒累積的器官，平均每個肺(右肺)將會有104 ~ 105的微粒累積。比較200 nm與800 nm的微粒累積，200 nm在各器官的平均微粒總累積量是800 nm的10倍左右。在24小時尿液當中，在第二週及第四週從部份800 nm暴露組小鼠尿液中檢測到塑膠微粒，在200 nm微粒暴露組則皆低於偵測極限。另外將肺部切片在螢光顯微鏡底下觀察，觀察到相比800 nm暴露組，200 nm微粒的暴露組肺臟累積的微粒含量要來得多，且大部分的微粒皆被巨噬細胞吞噬而聚集。 暴露組的肝臟重量顯著比控制組輕，且800 nm組別要比200 nm組別要來的輕，顯示較多的800 nm微粒累積在腸道將會導致菌群紊亂，肝臟重量下降。另外尿液中活性氧化物(ROS)為系統性氧化壓力，可以看到暴露組在8-OHdG、8-NO2Gua、8-IsoPF2a及HNE-MA的濃度均顯著比控制組要來得高(p<0.05)，且其中HNE-MA在200 nm微粒暴露組顯著比800 nm微粒暴露組來得高。此外，發炎因子的結果顯示在800 nm暴露組的在腎臟的IFNγ、IL-1β、IL-6與TNFα皆顯著高於控制組(p<0.05)，顯示800 nm微粒在腎臟排出的過程中可能造成了腎臟的發炎損傷，導致總排尿量顯著下降，肺臟發炎因子相較控制組則是沒有顯著變化。 上述實驗結果顯示相比800 nm微粒，200 nm微粒更能穿越腸胃道屏障累積在肝、肺、腎及脾臟當中，其中肺臟為主要累積的器官。部分800 nm微粒將會經由腎臟排泄進入尿液，造成腎臟發炎因子的上升，而200 nm微粒傾向累積在小鼠體內而不易被排出，並且兩者粒徑的微粒皆會引起全身性氧化壓力。本實驗在四週的重複暴露後，顯示較小的塑膠微粒較容易穿越腸胃道屏障並累積在哺乳類動物體內，產生氧化壓力及發炎反應，後續應繼續關注在長期重複暴露次微米或奈米塑膠微粒的毒性研究。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-25T03:05:12Z (GMT). No. of bitstreams: 1 U0001-0507202118251300.pdf: 12082930 bytes, checksum: 9ca8cb32083a04cce74491baa68af98c (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | 口試委員會審定書 i 誌謝 ii 中文摘要 iii 英文摘要 v 第一章 前言與研究目的 1 1.1. 前言 1 1.2. 研究目的 2 第二章 文獻回顧 3 2.1. 塑膠微粒的來源及分佈 3 2.2. 塑膠微粒的暴露途徑 5 2.3. 塑膠微粒的健康效應 7 2.3.1. 微粒與氧化壓力 7 2.3.2. 微粒與發炎因子 8 2.4. 塑膠微粒動力學 9 2.4.1. 腸胃道的吸收機制 9 2.4.2. 微粒在器官的分佈 11 2.4.3. 微粒的代謝與排泄 13 2.5. 塑膠微粒的檢測 14 第三章 材料與方法 15 3.1. 實驗流程與架構 15 3.2. 實驗動物 16 3.3. 聚苯乙烯塑膠微粒 17 3.3.1. 微粒的型態 17 3.3.2. 微粒的粒徑大小及表面電荷 17 3.3.3. 微粒的數目濃度 18 3.3.4. 微粒的螢光 18 3.3.5. 微粒懸浮液的製備 20 3.4. 暴露方法 22 3.5. 動物檢體收集與處理 23 3.5.1. 尿液收集 23 3.5.2. 動物犧牲與器官採樣 23 3.5.3. 腸道的取樣與前處理 24 3.5.4. 組織包埋 25 3.6. 組織中微粒的檢測 26 3.6.1. 組織消化液 26 3.6.1.1. 組織消化液的製備與檢測 26 3.6.1.2. 組織消化液的偵測極限 27 3.6.1.3. 消化對微粒的影響 28 3.6.1.4. 組織中微粒型態 29 3.6.2. 組織切片 29 3.6.2.1. 單一螢光影像觀察 29 3.6.2.2. 大圖螢光拼接觀察 30 3.7. 血液及尿液中微粒的檢測 31 3.7.1. 取樣與微粒的檢測 31 3.7.2. 血液與尿液中的偵測極限 32 3.7.3. 尿液中微粒型態 32 3.8. 生物指標 33 3.8.1. 尿液系統性氧化壓力 33 3.8.2. 特定器官發炎因子 34 3.9. 組織病理 36 3.10. 統計方法 36 第四章 結果 37 4.1. 實驗流程與架構 37 4.1.1. 體重變化 37 4.1.2. 飲水量與排尿量 37 4.1.3. 器官重量 38 4.2. 塑膠微粒的描述 39 4.2.1. 微粒的型態 39 4.2.2. 微粒的粒徑大小及表面電荷 39 4.2.3. 微粒的數目濃度 39 4.2.4. 微粒的螢光 40 4.3. 組織中微粒 41 4.3.1. 組織消化液的檢測 41 4.3.1.1. 微粒在腸道中的分佈 41 4.3.1.2. 微粒在器官的分佈 41 4.3.1.3. 微粒的偵測極限及回收率 41 4.3.1.4. 組織中微粒型態 42 4.3.2. 組織切片的觀察 43 4.3.2.1. 單一螢光影像觀察 43 4.3.2.2. 大圖螢光拼接觀察 43 4.4. 血液尿液中微粒 44 4.4.1. 微粒的檢測 44 4.4.2. 在血液與尿中液的偵測極限 45 4.4.3. 血尿中微粒型態 45 4.5. 生物指標 46 4.5.1. 尿液系統性氧化壓力 46 4.5.2. 特定器官發炎因子 46 4.6. 微粒分佈與生物指標相關性 47 4.6.1. 微粒在器官分佈的相關性 47 4.6.2. 微粒與氧化壓力的相關性 47 4.6.3. 微粒與發炎因子的相關性 48 4.7 組織病理 49 第五章 討論 50 5.1. 次微米塑膠微粒對動物生理指標的變化 51 5.2. 次微米塑膠微粒在腸道及器官的分佈 53 5.3. 次微米塑膠微粒與生物指標之關係 57 5.4. 研究限制 61 第六章 結論 63 第七章 建議 64 第八章 參考資料 65 | |
| dc.language.iso | zh-TW | |
| dc.subject | 發炎因子 | zh_TW |
| dc.subject | 次微米塑膠微粒 | zh_TW |
| dc.subject | 胃管灌食 | zh_TW |
| dc.subject | 小鼠 | zh_TW |
| dc.subject | 吸收效率 | zh_TW |
| dc.subject | 組織分佈 | zh_TW |
| dc.subject | 流式細胞儀 | zh_TW |
| dc.subject | 氧化壓力 | zh_TW |
| dc.subject | oral gavage | en |
| dc.subject | submicroplastics | en |
| dc.subject | cytokine | en |
| dc.subject | oxidative stress | en |
| dc.subject | flow cytometry | en |
| dc.subject | distribution | en |
| dc.subject | uptake | en |
| dc.subject | mice | en |
| dc.title | 次微米塑膠微粒在小鼠經由腸胃道的吸收分佈及毒性研究 | zh_TW |
| dc.title | The distribution and toxicity of submicron plastic particles through the gastrointestinal tract in mice | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 姜至剛(Hsin-Tsai Liu),蕭伊倫(Chih-Yang Tseng),吳焜裕 | |
| dc.subject.keyword | 次微米塑膠微粒,胃管灌食,小鼠,吸收效率,組織分佈,流式細胞儀,氧化壓力,發炎因子, | zh_TW |
| dc.subject.keyword | submicroplastics,oral gavage,mice,uptake,distribution,flow cytometry,oxidative stress,cytokine, | en |
| dc.relation.page | 133 | |
| dc.identifier.doi | 10.6342/NTU202101281 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2021-07-12 | |
| dc.contributor.author-college | 公共衛生學院 | zh_TW |
| dc.contributor.author-dept | 環境與職業健康科學研究所 | zh_TW |
| dc.date.embargo-lift | 2023-01-01 | - |
| 顯示於系所單位： | 環境與職業健康科學研究所 | |
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| U0001-0507202118251300.pdf | 11.8 MB | Adobe PDF | 檢視/開啟 |
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