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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 侯詠德(Yung-Te Hou) | |
| dc.contributor.author | Yu-Che Lin | en |
| dc.contributor.author | 林于哲 | zh_TW |
| dc.date.accessioned | 2022-11-24T03:20:36Z | - |
| dc.date.available | 2021-10-04 | |
| dc.date.available | 2022-11-24T03:20:36Z | - |
| dc.date.copyright | 2021-10-04 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-09-27 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/80888 | - |
| dc.description.abstract | 肝臟是人體重要的生化工廠，具有許多不可取代之功能。但目前肝衰竭 (Liver failure) 及肝硬化 (Liver cirrhosis) 等疾病之治療手段由於受限於患者本身肝臟機能之差異而有不同程度之處理方式，因此提升肝臟機能以利重症患者之手術能成功執行也成為目前亟需解決之問題。傳統中草藥 (Traditional Chinese herbal medicine) 用於醫學治療及健康保健已有數千年之歷史，而將現有之中草藥開發於新型態肝臟治療之研究也有其應用潛能。本研究擬透過天然植物多酚之單寧酸 (Tannic acid; TA) 來闡明其對肝細胞活性及機能提升的影響，並接著使用體外及體內毒化衰竭模型來進一步確認單寧酸對於肝細胞機能與活性之影響的機制。本研究結果可知：(1) 增強肝機能的最佳 TA 濃度約為 0.5 至 5 μg/mL；(2) TA 對初代成熟肝細胞的體外培養有明顯的短期機能提升之效果 (培養初代成熟肝細胞第 3 天後其相對增值率增加 10%；培養從第 2 天到第 3 天時白蛋白合成增加 18%；培養第 2 天時白蛋白的基因表達增加 210%)；(3) TA 對小型肝細胞的體外培養亦有長期且明顯的機能提升之效果 (小型肝細胞培養第 7 天後其相對增值率增加 7%；培養從第 1 天到第 11 天時白蛋白合成增加至 300%；培養第 1 天至第 11 天時白蛋白的基因表達增加約 200% 至 400%)；(4) TA 能修復經過四氯化碳 (Carbon tetrachloride; CCl4) 及乙醯胺酚 (Acetaminophen; APAP) 所毒化的初代成熟肝細胞 (將以 CCl4 預毒化處理後之初帶成熟肝細胞於含有 TA 之培養液回復 4 至 8 小時，其與未加入 TA 回復之肝細胞毒性相比降低 40% 至 90%；將以 APAP 預毒化處理後之肝細胞於含有 TA 之培養液回復 18 小時，其與未加入 TA 回復之初代成熟肝細胞及小型肝細胞的氧化壓力下降約 10% 至 33%)。 本項研究顯示，TA 在特定的添加濃度下具有增強肝細胞機能的潛力。此外，肝細胞在藉由含 APAP/CCl4 培養液 (用於模擬體外毒素介導的肝纖維化/急性肝衰竭) 造成損傷後，不論是在體內/體外實驗都有機會在另外添加 TA 之後回復到其損傷前狀態之潛能。因此本研究對於「喝茶保肝」之說完成科學之認證，並有望進一步應用於臨床醫學上。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-24T03:20:36Z (GMT). No. of bitstreams: 1 U0001-2309202112063500.pdf: 5744049 bytes, checksum: 858c2e3268ce6a4ef02fef989f92f1d8 (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | 誌謝 i 摘要 iii Abstract v 目錄 vii 圖目錄 xi 表目錄 xvi 第一章 前言 1 1.1 研究背景與目的 1 1.2 實驗架構 3 第二章 文獻探討 5 2.1 肝病治療發展近況 5 2.1.1 肝衰竭 (Liver failure) 5 2.1.2 肝纖維化 (Liver fibrosis) 7 2.1.3 肝臟移植 (Liver transplantation) 8 2.1.4 肝臟組織再生 (Liver tissue regeneration) 9 2.2 細胞來源 10 2.2.1 成熟肝細胞 (Mature hepatocytes) 10 2.2.2 小型肝細胞 (Small hepatocytes) 11 2.3 材料性質與介紹 13 2.3.1 單寧酸 (Tannic acid) 13 2.4 本研究欲達成之目標 (Specific aims) 16 2.4.1 單寧酸對肝細胞之體外機能促進探討 16 2.4.2 將單寧酸應用於細胞層級之治療開發 16 2.4.3 將單寧酸應用於體內急性肝衰竭之治療 16 第三章 研究方法 18 3.1 實驗藥品、耗材、儀器設備與實驗動物 18 3.1.1 實驗藥品 18 3.1.2 實驗耗材 20 3.1.3 實驗儀器設備 21 3.1.4 實驗動物 22 3.2 肝細胞採取與培養 23 3.3 實驗相關檢測方法 25 3.3.1粒線體活性檢測 (MTT Assay) 25 3.3.2相對細胞生存率檢測 26 3.3.3肝蛋白 (Albumin) 分泌量檢測 27 3.3.4乳酸脫氫酵素 (LDH) 活性檢測 28 3.3.5尿素 (Urea) 活性檢測 29 3.3.6反轉錄即時定量PCR (Quantitative real-time PCR) 30 3.3.7氧化壓力 (Oxidation stress) 檢測 33 3.3.8以流式細胞儀進行細胞凋亡 (Apoptosis) 之檢測 34 3.4 體外毒化實驗處理 35 3.4.1 四氯化碳毒化 35 3.4.2 乙醯胺酚毒化 36 3.5 體內實驗分析相關 39 3.5.1血清生化分析 39 3.5.2組織切片分析 40 第四章 結果與討論 41 4.1 單寧酸對體外培養初代成熟肝細胞之影響 41 4.1.1 單寧酸對初代成熟肝細胞培養之增殖能力的影響 41 4.1.2 單寧酸對初代成熟肝細胞培養之細胞生存率的影響 45 4.1.3 單寧酸對初代成熟肝細胞培養之白蛋白分泌量的影響 47 4.1.4 單寧酸對初代成熟肝細胞培養之尿素分泌量的影響 49 4.1.5 單寧酸對初代成熟肝細胞培養之機能基因表現量的影響 51 4.1.6 單寧酸對初代成熟肝細胞培養之細胞毒性的影響 53 4.1.7 單寧酸對初代成熟肝細胞培養之氧化壓力的影響 54 4.1.8 單寧酸對初代成熟肝細胞培養對細胞凋亡的影響 57 4.1.9 單寧酸對初代成熟肝細胞培養之生長型態比較 60 4.2 單寧酸對體外初代成熟肝細胞培養毒化後回復之影響 62 4.2.1 單寧酸對初代成熟肝細胞於 CCl4 毒化後回復之增殖能力的影響 62 4.2.2 單寧酸對初代成熟肝細胞於 CCl4 毒化後回復之尿素分泌量的影響 66 4.2.3 單寧酸對初代成熟肝細胞於 CCl4 毒化後回復之細胞毒性的影響 68 4.2.4 單寧酸對初代成熟肝細胞於 APAP 毒化後回復之增殖能力的影響 70 4.2.5 單寧酸對初代成熟肝細胞於 APAP 毒化後回復之白蛋白分泌量的影響 73 4.2.6 單寧酸對初代成熟肝細胞於 APAP 毒化後回復之尿素分泌量的影響 75 4.2.7 單寧酸對初代成熟肝細胞於 APAP 毒化後回復之基因表現量的影響 78 4.2.8 單寧酸對初代成熟肝細胞於 APAP 毒化後回復之細胞毒性的影響 81 4.2.9 單寧酸對初代成熟肝細胞於 APAP 毒化後回復之氧化壓力的影響 84 4.2.10單寧酸對初代成熟肝細胞於 APAP 毒化後回復對細胞凋亡的影響 87 4.2.11 單寧酸對初代成熟肝細胞於 APAP 毒化後回復生長型態之比較 90 4.3 單寧酸對體外培養小型肝細胞之影響 92 4.3.1 單寧酸對小型肝細胞培養之增殖能力的影響 92 4.3.2 單寧酸對小型肝細胞培養之白蛋白分泌量的影響 94 4.3.3 單寧酸對小型肝細胞培養之尿素分泌量的影響 96 4.3.4 單寧酸對小型肝細胞培養之機能基因表現量的影響 98 4.3.5 單寧酸對小型肝細胞培養之細胞毒性的影響 100 4.3.6 單寧酸對小型肝細胞培養之氧化壓力的影響 102 4.3.7 單寧酸對小型肝細胞培養對細胞凋亡的影響 104 4.3.8 單寧酸對小型肝細胞培養之生長型態比較 107 4.4 單寧酸對體外培養小型肝細胞毒化後回復之影響 109 4.4.1 單寧酸對小型肝細胞於 APAP 毒化後回復之增殖能力的影響 109 4.4.2 單寧酸對小型肝細胞於 APAP 毒化後回復之細胞生存率的影響 113 4.4.3 單寧酸對小型肝細胞於 APAP 毒化後回復之白蛋白分泌量的影響 115 4.4.4 單寧酸對小型肝細胞於 APAP 毒化後回復之尿素分泌量的影響 117 4.4.5 單寧酸對小型肝細胞於 APAP 毒化後回復之細胞毒性的影響 120 4.4.6 單寧酸對小型肝細胞於 APAP 毒化後回復之氧化壓力的影響 123 4.4.7 單寧酸對小型肝細胞於 APAP 毒化後回復培養之細胞凋亡的影響 126 4.5 單寧酸對體內肝細胞預防及毒化後回復之效果影響 129 4.5.1 血清生化分析結果 129 4.5.2組織切片分析結果 134 第五章 結論與未來展望 138 5.1結論 138 5.2未來展望 141 參考文獻 145 | |
| dc.language.iso | zh-TW | |
| dc.subject | 急性肝衰竭 | zh_TW |
| dc.subject | 單寧酸 | zh_TW |
| dc.subject | 肝組織工程 | zh_TW |
| dc.subject | 肝細胞 | zh_TW |
| dc.subject | Hepatocyte | en |
| dc.subject | APAP-induced liver injury | en |
| dc.subject | Liver tissue engineering | en |
| dc.subject | Tannic acid | en |
| dc.title | 單寧酸對於肝臟保肝效果及肝臟機能提升效果之研究 | zh_TW |
| dc.title | The Effects of Tannic Acid on Ameliorating Hepatic Functions and Hepatoprotective Ability | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 林峰輝(Hsin-Tsai Liu),倪衍玄(Chih-Yang Tseng) | |
| dc.subject.keyword | 單寧酸,肝組織工程,肝細胞,急性肝衰竭, | zh_TW |
| dc.subject.keyword | Tannic acid,Hepatocyte,Liver tissue engineering,APAP-induced liver injury, | en |
| dc.relation.page | 151 | |
| dc.identifier.doi | 10.6342/NTU202103312 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2021-09-28 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 生物機電工程學系 | zh_TW |
| 顯示於系所單位： | 生物機電工程學系 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| U0001-2309202112063500.pdf 授權僅限NTU校內IP使用（校園外請利用VPN校外連線服務） | 5.61 MB | Adobe PDF |
系統中的文件，除了特別指名其著作權條款之外，均受到著作權保護，並且保留所有的權利。