臺灣博碩士論文加值系統

近年來，隨著健康意識抬頭，空氣污染的問題逐漸受到重視，環境中PM2對公眾健康造成的影響已經成為全球人們關注的焦點，近期多個研究指出，肺癌的發生率提高和PM2有著密不可分的關係。在台灣，癌症一直是十大死因之首，且以肺癌的致死率最高，這可能與空氣汙染中的PM2有關。現代社會，人們越來越注重養生，從天然來源萃取出的多醣日益受到關注，台灣四面環海，海洋資源豐富，本研究所使用的海木耳屬於紅藻的一種，利用熱水萃取的方式萃得海木耳水萃物，實驗動物為C57BL/6，將小鼠進行分組後，腫瘤背負組皮下注射小鼠肺癌細胞 (LLC)，再以噴霧的方式讓各兩的組正常小鼠和腫瘤背負小鼠暴露於PM2的環境中，且其中一組餵食海木耳多醣，探討海木耳水萃物抗腫瘤的效果，結果顯示，海木耳水萃物具有抗氧化及抗發炎的能力，在細胞實驗中也觀察到其具有抑制癌細胞生長、遷移和侵襲的能力，動物實驗結果顯示，海木耳水萃物可輔助改善腫瘤所引起的惡病質症狀，也能抑制腫瘤的轉移及生長。後續分析發現海木耳水萃物會增加毒殺型 T 細胞 (Cytotoxic T cell) 及自然殺手細胞 (NK cell) 表現亮，經 ELISA 測定小鼠血清中 IFN-γ 表現量，發現餵食海木耳水萃物能使小鼠的 IFN-γ 表現量上升，因此推測海木耳水萃物是藉由活化 Th1 路徑來達到毒殺癌細胞的效果。並提升免疫細胞數量，達到毒殺癌細胞之效果。

Growing health consciousness in recent years has led people to pay more attention to air pollution, The impacts of ambient PM2 on public health had become great concerns worldwide. Recently, more and more studies indicate that PM2 has been associated with an increase in lung cancer incidence. In Taiwan, cancer is the first among the top ten leading causes of death, and lung cancer has the highest fatality rate. Taiwan is surrounded by sea, and have abundant ocean sources. Sarcodia montagneana is a kind of red seaweed, in this study we used Sarcodia montagneana water extracts to fed C57BL/6 mice. During the experiment, we use an atomizer to spray PM2, made normal mice and tumor-bearing mice exposed to PM2. The goal of the study is to evaluate the anti-tumor effect of Sarcodia montagneana water extracts. In vitro, Sarcodia montagneana water extracts could inhibit LLC cells on migration and invasion in transwell assay. In vivo, Sarcodia montagneana water extracts could increase the immune cell, decrease body-weight loss and tumor weight, through ELISA kit analysis, we found that the group who have been fed Sarcodia montagneana water extracts, increased the level of IFN-γ in serum. In conclusion, Sarcodia montagneana water extracts could anti-tumor growth, cancer cachexia and inhibit metastatic activities through Th1 pathway.

摘要 I
Abstract II
目錄 III
壹、前言 1
貳、文獻回顧 3
一、癌症 3
二、肺癌 (Lung cancer) 3
1. 肺癌病理學 3
2. 肺癌流行病學 3
3. 肺癌特徵 4
4. 肺癌的治療與困境 4
二、懸浮微粒 (Particulate matter, PM) 4
1. 空氣中的懸浮微粒 4
2. 懸浮微粒與疾病之間的關係 5
三、藻類 5
1. 海藻(Seaweed) 5
1.1海藻之簡介 5
1.2台灣海藻的分布 6
1.3紅藻 (Red algae) 6
2. 海木耳 (Sarcodia montagneana) 6
2.1海木耳之簡介 6
2.2海木耳生理活性 7
參、實驗設計流程 8
一、海木耳萃取及特性分析 8
二、細胞實驗 (In vitro) 8
三、動物實驗 (In vivo ) 9
肆、材料與方法 10
一、實驗材料 10
1. 實驗細胞株 10
2. 細胞培養關試劑 10
3. 海木耳來源 10
4. PM2來源 10
5. 細胞存活率試劑 10
6. 抗氧化試驗 10
7. 動物實驗 11
8. 細胞激素分析 11
9. 蛋白質萃取與西方墨點法相關試劑 11
10. 組織切片 12
11. 免疫螢光染色 12
12. 免疫化學染色 12
二、實驗方法 13
1. 細胞株之保存活化與培養 13
1.1 細胞株之保存 13
1.2 細胞株活化 13
1.3 細胞培養 13
2. 海木耳樣品製備 13
2.1 熱水萃取法 14
3. PM2收集 14
4. 樣品特性分析 14
4.2 總醣含量測定 (Total sugar content) 14
4.3 硫酸根含量測定 15
4.4 海木耳對細胞毒性測試 15
4.5 抗氧化能力試驗 15
4.6 細胞內一氧化氮含量測定 17
5.細胞遷移試驗 17
5.1 傷口癒合試驗 17
5.2細胞遷移和侵襲試驗 17
5.3 細胞侵襲試驗 18
6. 動物實驗 (In vivo) 18
6.1 實驗小鼠模式建立 18
6.2 組織器官分析 18
7. 統計分析 19
伍、結果 20
一、海木耳水萃物特性分析 20
二、海木耳水萃物抗氧化活性之探討 20
三、體外試驗 20
1. 海木耳水萃物對細胞毒性之影響 20
2. PM2對細胞毒性之影響 20
3. 海木耳水萃物對巨噬細胞吞噬能力之影響 20
4. PM2對巨噬細胞吞噬能力之影響 21
5. 傷口癒合試驗 21
6. 細胞遷移及侵襲試驗 21
四、利用動物實驗探討海木耳水萃物及PM2對腫瘤背負小鼠之影響 22
1. 海木耳水萃物及PM2對腫瘤生長之影響 22
2. 海木耳水萃物及PM2對肺癌腫瘤背負小鼠體重變化之影響 22
3. 海木耳水萃物及PM2對肌肉及脂肪之影響 22
4. 海木耳水萃物及PM2對腫瘤組織中 COX-2 蛋白表現量之影響 22
5. 海木耳水萃物抑制腫瘤轉移 22
6. 海木耳水萃物對淋巴細胞之調節作用 23
7. 海木耳水萃物對Th1及Th2之調節作用 23
8. 海木耳水萃物對腫瘤血管新生的影響 23
陸、討論 24
柒、結論 28
捌、參考文獻 29
玖、圖表 36


 

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