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研究生:梁安
研究生(外文):An Liang
論文名稱:飲食中不同n-6與n-3脂肪酸比例及蝦紅素對於誘發性口腔癌症之影響
論文名稱(外文):Effects of Various Dietary n-6/n-3 Fatty Acids and Astaxanthin on Induced Oral Carcinoma
指導教授:黃士懿黃士懿引用關係
口試委員:林永和楊素卿
口試日期:2015-06-26
學位類別:碩士
校院名稱:臺北醫學大學
系所名稱:保健營養學研究所
學門:醫藥衛生學門
學類:營養學類
論文種類:學術論文
論文出版年:2015
畢業學年度:103
語文別:中文
論文頁數:149
中文關鍵詞:口腔癌檳榔嚼塊n-6/n-3脂肪酸比例蝦紅素腫瘤負荷
外文關鍵詞:oral carcinomabetel quiddietary fatty acidsastaxanthintumor burden
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根據流病統計的結果發現,口腔癌的發生率日益上升,且主要發生在台灣、印度等有嚼食檳榔嚼塊習慣的東南亞國家。嚼食檳榔時,會因檳榔嚼塊中之化學成分及其粗纖維造成口腔黏膜發炎並增加其氧化壓力,造成損傷。長期嚼食檳榔將導致口腔癌症之發生。過去的研究指出,降低發炎反應與氧化壓力皆可能是預防或是治療癌症的方式。文獻指出,飲食中的脂肪酸組成可能與某些特定癌症的發生有關,改變飲食中脂肪酸之組成可能可以降低特定癌症之發生率;另有文獻顯示,類胡蘿蔔素具有強大的清除自由基之能力,並可調控細胞週期達到癌症化學預防之功效。故本研究利用7,12-dimethylbenz[a]anthracene(DMBA)及檳榔嚼塊萃取物來誘發倉鼠口腔癌前病變,透過調整實驗飼料中n-6及n-3脂肪酸的比例且在不同的n-6/n-3飲食下以塗抹的方式給予蝦紅素,以探討是否能夠降低實驗動物體內發炎反應與氧化壓力,達到預防或是改善口腔癌之發生。研究設計使用45隻6週齡之雄性倉鼠,依照實驗飼料的不同分為正常(N組,n-6/n-3=6)、低(L組,n-6/n-3=2)與高(H組,n-6/n-3=15) n-6/n-3脂肪酸飲食,並依照不同的塗抹物質各自再細分為正常組(N)、負控制組(C)及蝦紅素組(A)。除了正常組(NN、LN及HN)外,其餘的六組(NC、LC、HC、NA、LA及HA)於實驗第1週至第5週會使用0.5% DMBA(溶於礦物油中)塗抹於倉鼠右側口腔頰囊,以誘發其口腔癌化,在實驗第6至16週時則以檳榔嚼塊萃取物(betel quid extract, BQE)取代0.5% DMBA進行塗抹。蝦紅素組(NA、LA及HA)則在實驗第6至16週時除了塗抹BQE外,同時以0.5%蝦紅素(溶於礦物油中)與BQE隔日交互塗抹於右側口腔頰囊。結果發現,給予倉鼠低n-6/n-3脂肪酸飲食(n-6/n-3=2)可以降低其體內n-6/n-3脂肪酸比例,向下調節口腔頰囊NF-κB、cyclin D1之表現量,並減少細胞增生指標─細胞增殖核抗原(proliferating cell nuclear antigen, PCNA)之表現量,達到顯著減少口腔腫瘤數目、腫瘤體積及腫瘤負荷。然而,無論在何種n-6/n-3脂肪酸飲食下,輔以塗抹0.5%之蝦紅素於具口腔癌前病變之倉鼠,均無法有效減緩其口腔癌前病變之發生。
Betel quid (BQ) chewing is thought to contribute to leukoplakia, submucosal fibrosis and oral cancer in Taiwan, India and many Southeast Asian countries. Dietary fatty acid patterns have been linked to certain cancer prevalence. Carotenoids (i.e. astaxanthin) have been indicated as possessing a strong ability to protect cells against active oxygen species by interfering with protein expression in the cell cycle. Thus, we investigated the chemopreventive effects of various dietary n-6/n-3 fatty acid and astaxanthin under DMBA/Betel quid extract (BQE)-induced hamster oral cancer model. Forty-five 6-week-old adult male hamsters were housed and divided into normal (N), low (L) and high (H) dietary n-6/n-3 fatty acid groups and subdiveid into normal (N), control (C) and astaxanthin (A) groups (n=5 for each subgroup). All animals except normal group were initially daubed with 0.2 mL of 0.5% DMBA solution to the right buccal pouch of animals for 5 weeks. In the subsequent 11 weeks, these hamsters were daubed with either BQE or a mineral oil solution 0.2 mL of 0.5% astaxanthin three times per week on alternate days. Results indicate the low dietary n-6/n-3 fatty acid group presented the significant lower tumor number, volume and tumor burden than other groups. Furthermore, the low dietary n-6/n-3 fatty acid group presented the significant lower NF-κB, cyclinD1 and PCNA expression in the right buccal pouch tissue and lower n-6/n-3 fatty acids profile in red blood cell than other groups. In conclusion, lower dietary n-6/n-3 fatty acids showed the chemopreventive effects on DMBA/BQE-induced hamster oral cancer model. However, our results indicated astaxanthin lack of chemopreventive effects under various n-6/n-3 fatty acids diets on DMBA/BQE-induced hamster oral cancer model.
中文摘要 I
英文摘要 III
致謝 V
目錄 X
表目錄 XVI
圖目錄 XVII
縮寫表 XIX
第一章 緒論 1
第二章 文獻回顧 3
第一節 口腔癌症 3
一、口腔癌症流行病學 3
二、口腔癌症簡介 6
三、口腔癌症之診斷、分期與治療 8
第二節 檳榔嚼塊組成與口腔癌之發生 10
第三節 口腔癌症之動物模式 13
一、倉鼠口腔頰囊之口腔癌症動物模式 13
二、4-Nitroquinoline 1-oxide誘發之口腔癌症模式 14
第四節 口腔癌症與發炎反應 15
第五節 飲食中脂肪酸組成與發炎反應 17
第六節 飲食中脂肪酸之組成與癌症 20
第七節 口腔癌症與氧化壓力 22
第八節 蝦紅素與口腔癌症 25
第九節 口腔癌化過程之生物標的 29
第三章 研究動機及目的 31
第四章 實驗設計與方法 31
第一節 實驗流程 31
第二節 實驗動物 34
第三節 口腔癌症之誘發 35
第四節 介入物質 37
一、實驗飼料油脂 37
二、蝦紅素 40
第五節 飼料組成 41
第六節 樣本收集與處理 43
一、飲食及體重紀錄 43
二、血液樣本 43
三、組織及臟器採集 43
第七節 分析項目與方法 45
一、血漿生化分析 45
二、實驗飼料油脂、血漿、紅血球脂肪酸組成 51
三、口腔頰囊腫瘤數目、體積及腫瘤負荷計算與病理切片分析 57
四、食道切片分析 63
五、肝臟及腎臟切片分析 63
第八節 統計分析 64
第五章 結果 65
第一節 實驗動物體重、肝臟及腎臟重量 65
一、實驗動物體重 65
第二節 口腔腫瘤數目、體積與腫瘤負荷 67
一、正常組與負控制組 67
二、正常組與蝦紅素組 67
第三節 實驗動物血漿脂肪酸分析 68
一、飽和脂肪酸 68
二、單元不飽和脂肪酸 68
三、n-6多元不飽和脂肪酸 68
四、n-3多元不飽和脂肪酸 69
五、n-6及n-3多元不飽和脂肪酸比值 69
第四節 實驗動物紅血球脂肪酸分析 70
一、飽和脂肪酸 70
二、單元不飽和脂肪酸 70
三、n-6多元不飽和脂肪酸 70
四、n-3多元不飽和脂肪酸 71
五、n-6及n-3多元不飽和脂肪酸比值 71
第五節 腫瘤體積、腫瘤負荷及紅血球脂肪酸組成之相關性 72
一、平均腫瘤體積大小與紅血球脂肪酸組成之相關性 72
二、腫瘤負荷與紅血球脂肪酸組成之相關性 72
第六節 血漿發炎反應細胞激素之比較 74
一、 腫瘤壞死因子-α 74
二、 介白素-6 74
第七節 口腔囊頰組織與細胞型態變化 75
一、 正常組 75
二、 負控制組 75
三、 蝦紅素組 75
第八節 口腔頰囊免疫組織化學染色 77
一、 NF-κB p65 77
二、 增殖細胞核抗原 77
三、 Cyclin D1 77
第九節 食道組織與細胞型態變化 78
一、 正常組 78
二、 負控制組 78
三、 蝦紅素組 78
第十節 肝臟及腎臟組織與細胞型態變化 79
一、 正常組 79
二、 負控制組 79
三、 蝦紅素組 79
第六章 討論 80
第一節 實驗動物體重 80
一、負控制組 80
二、蝦紅素組 80
第二節 實驗動物肝臟及腎臟重量與組織變化 81
一、負控制組 81
二、蝦紅素組 81
第三節 飲食介入對於誘發性口腔癌症之巨觀影響 82
一、不同n-6/n-3脂肪酸飲食對於口腔腫瘤數目、體積及腫瘤負荷之影響 82
二、蝦紅素於不同n-6/n-3脂肪酸飲食下對於口腔腫瘤數目、體積及腫瘤負荷之影響 83
第四節 飲食介入對於脂肪酸組成之影響 86
第五節 飲食介入對於血漿中發炎細胞激素之影響 87
第六節 飲食介入對於誘發性口腔癌症之微觀影響 88
第七節 飲食介入對於食道組織之影響 91
第八節 綜合討論 92
第七章 結論 93
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