臺灣博碩士論文加值系統

5-氟尿嘧啶 (5-FU) 是治療癌症的藥物之一；褐藻醣膠具有調節免疫細胞的功能。本研究使用幾丁聚醣/褐藻醣膠功能性奈米粒，包覆 5-FU，期望結合免疫治療和化學療法來治療癌症。以幾丁聚醣與褐藻醣膠重量比 = 5：1 製備奈米粒，以幾丁聚醣/褐藻醣膠奈米粒包覆 5-氟尿嘧啶，奈米粒粒徑為 151.19±9.06 nm、表面電位為 28.11±3.01 mV 以及 PDI 值為 0.22±0.03；使用 HPLC 檢測到包覆率為 73.84±0.83 ％、裝載量為 5.03±0.05 ％。將幾丁聚醣/褐藻醣膠聚合物與 5-FU 的比例改變，進行體外藥物釋放實驗，結果顯示：聚合物比例變大，則藥物的釋放量較少、釋放較緩慢；藥物比例增大，突釋較快，無法保持長時間釋放。細胞實驗結果顯示，包覆 5-FU 之奈米粒對人類結腸癌細胞株 (HT-29) 有較強的毒殺效果、顯著增加癌細胞的凋亡程度，透過雷射共軛焦掃描電子顯微鏡亦可觀察到奈米粒被 HT-29 細胞攝取；然而此奈米粒對小鼠纖維母細胞 (L929) 並無增強其毒殺效果。此外，包覆 5-FU之奈米粒亦能誘導巨噬細胞 (RAW264.7) 分泌 TNF-α、GM-CSF，進一步調節免疫系統活化。根據上述實驗結果顯示包覆 5-FU 之幾丁聚醣/褐藻醣膠奈米粒系統可經化學和免疫活化來治療癌症，為有潛力的治療癌症藥劑。

5-Fluorouracil (5-FU) is widely used in the treatment of cancer, and fucoidan can function as an adjuvant to regulate immune cells such as enhancing cell maturation and inducing immune responses. In this study, we developed 5-FU coated chitosan/fucoidan (CS/F) functional nanoparticles (NPs), which are expected as a desired treatment to combine immunotherapy with chemotherapy for cancer treatment. As the CS to F weight ratio was 5:1, the NP shows the best performance. Its diameter was 151.19 ± 9.06 nm, the zeta potential was 28.11 ± 3.01 mV and PDI was 0.22 ± 0.03. The encapsulation efficiency of 5-FU in CS/F NPs was 73.84 ± 0.83%, the loading capacity was 5.03 ± 0.05%. We observe in vitro drug release by modifying the weight ratio of polymer (CS and F) to 5-FU. As the polymer ratio increases, the release rate and amount of 5-FU are decreased; as 5-FU ratio increases, 5-FU is burst released from NPs and unable to maintain a long time. The 5-FU coated CF/S NPs have a strong toxic effect on colon carcinoma cell line HT-29 cells and can inducing HT-29 cells apoptosis. Furthermore, HT-29 cellular uptake of NPs was observed by confocal laser scanning transmission electron microscope. However, The 5-FU coated CF/S NPs showed no toxic effects on mouse fibroblasts cell line L929. In addition, the 5-FU coated CF/S NPs can induce TNF-α and GM-CSF secretion of macrophage (RAW264.7), and regulate the immune response promptly. According to the results, the 5-FU coated CF/S NPs are potential cancer therapy agents and expected as a desired treatment combining immunotherapy with chemotherapy to treat cancer.

謝誌 I
摘要 II
Abstract III
目錄 IV
圖目錄 VI
表目錄 VIII
第一章、前言 1
第二章、文獻回顧 2
2.1 癌症 2
2.1-1 癌症的成因 2
2.1-2 癌症症狀 2
2.1-3 癌症治療 2
2.2 5-氟尿嘧啶 3
2.2-1 5-氟尿嘧啶的用途 3
2.2-2 5-氟尿嘧啶的作用機制 3
2.2-3 5-氟尿嘧啶的副作用 4
2.3 褐藻醣膠 4
2.3-1 褐藻醣膠結構 4
2.3-2 褐藻醣膠的抗癌活性 4
2.3-3 褐藻醣膠的免疫調節能力 4
2.3-4 褐藻醣膠奈米粒 5
2.4 幾丁聚醣/褐藻醣膠奈米粒 5
第三章、實驗目的 7
第四章、實驗架構 8
第五章、實驗流程 9
第六章、實驗材料與方法 10
6.1實驗材料 10
6.1.1實驗藥品 10
6.1.2實驗儀器 11
6.1.3實驗溶液配製 13
6.2實驗方法 14
6.2.1 幾丁聚醣的製備 14
6.2.2 褐藻醣膠成分分析 14
6.2.3 製備幾丁聚醣/褐藻醣膠奈米粒 15
6.2.4 製備幾丁聚醣/褐藻醣膠/5-氟尿嘧啶奈米粒 15
6.2.5 幾丁聚醣/褐藻醣膠奈米粒之物理化學特性探討 15
6.2.6 幾丁聚醣/褐藻醣膠/5-氟尿嘧啶奈米粒之物理化學特性探討 16
6.2.7 5-氟尿嘧啶之釋放分析 17
6.2.8 奈米粒的抗癌效果 17
6.2.9 奈米粒的免疫調節 18
6.2.10 統計分析 19
第七章、結果討論 20
7.1 幾丁聚醣的製備與褐藻醣膠的成分 20
7.2 材料在不同 pH 值下的電荷密度 20
7.3 幾丁聚醣/褐藻醣膠/5-氟尿嘧啶奈米粒之物理化學特性 20
7.4 控制釋放 22
7.5 奈米粒構形觀察 22
7.6 細胞實驗 22
第九章、參考文獻 27
第十章、圖表 32

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