臺灣博碩士論文加值系統

本實驗室過去的研究已證實，給予懷孕期母鼠攝取21天的高脂飲食，其子代罹患乳腺腫瘤的比例與子代在出生後攝取230天之高脂飲食而罹患乳腺腫瘤的比例相當，顯示懷孕期的飲食對於日後子代罹患乳癌的風險有很重要的影響。且在過去文獻指出，除了estradiol (E2) 以外，其代謝產物 (estrogen metabolites) 亦與乳癌的發生有密切相關。因此本論文主要對於生殖期 (reproductive age) 與懷孕期 (pregnancy) 之雌鼠，給予攝取高n-6脂肪酸飲食，探討飲食中脂肪酸的組成與脂肪含量的高低對尿液雌激素代謝產物之影響。實驗組別處理主要可分為三大組，分別為生殖期雌鼠給予高n-6脂肪酸飲食，並於餵食80天與120天後收集尿液作分析、 懷孕期雌鼠給予高n-9脂肪酸或高n-6脂肪酸飲食配合魚油補充，分別於懷孕第10天與第18天收集尿液作分析以及懷孕期雌鼠給予低脂與高脂飲食配合魚油補充，分別於第10天與第18天收集尿液作分析。在建立較適合的雌激素代謝產物純化方法進行純化後，以HPLC配合8個電極之電化學偵測器(ECD) 進行檢測。實驗結果顯示，給予生殖期或懷孕期母鼠攝取高n-6脂肪酸，比起給予chow diet飲食，明顯增加尿液中total estrogen metabolites、16α-OHE1、E3與2-OHE1之含量，其中又以2-OHE1增加最多，且易降低2-OCH3E/ 2-OHE與4-OCH3E/ 4-OHE，顯示高n-6脂肪酸會抑制hydroxyestrogen的甲基化。此外，給予懷孕期母鼠攝取18天高n-9脂肪酸飲食後，與高n-6脂肪酸飲食有相似的作用，易提高罹患乳癌的風險。由以上結果可知，高n-6脂肪酸會透過影響雌激素的代謝而造成罹患乳癌風險的提高。

Breast cancer is associated with high-fat intake. About two thirds of breast cancer in women are estrogen-dependent that contain estrogen receptor . It has been found that not only estrogen, the estrogen metabolites can also bind with estrogen receptor for the regulation of breast cancer development. We aimed to examine the effect of high fat diet on estrogen metabolism during reproductive age and pregnancy in rats. The urine was collected during 5-day estrus cycle in adult rats fed chow or high n-6 sunflower oil diets for 80 days, or collected at gestation day 10 or day 18 in pregnant rats fed the chow diet, high n-9 safflower oil, high n-6 sunflower oil without or with fish oil supplementation, low fat or high fat diets without or with fish oil supplementation. The 12 estrogen metabolites in urine was measured by HPLC with 8-channel electrochemical detector. We found that total estrogen metabolites, 16 -hydroxyestrone, 2-hydroxyestrone and estriol were increased in rat urine fed the high n-6 sunflower oil diet during reproductive age and in pregnancy compared with that fed chow or low fat diets. The methylation of 4-hydroxyestrogen and 2-hydroxyestrogen was decreased in rat urine fed the high n-6 sunflower oil or high n-9 safflower oil diets during reproductive age and in pregnancy compared with that
fed chow or low fat diets. And this decreased effect of methylation of 2-hydroxyestrogen by high n-6 fat diet was increased back by fish oil supplementation during pregnancy. We proposed that carcinogen 16 -hydroxyestrone is increased and the anti-carcinogen 2-methoxyestrone is decreased in female rats fed high fat diet.

中文摘要 I
Abstract II
圖目錄 VI
第一章、文獻回顧 1
一、乳癌 1
二、飲食與乳癌 1
三、油脂與代謝 3
四、雌激素與乳癌 5
五、雌激素代謝產物與乳癌 6
第二章、實驗目的與假設 10
一、實驗目的 10
1. Rat urinary estrogen metabolites萃取前處理方法的建立 10
2. Urinary estrogen metabolites分析 10
二、實驗假設 11
第三章、材料與方法 12
一、尿液樣品收集 12
二、尿液雌激素代謝產物萃取 12
1. 尿液雌激素代謝產物前處理與萃取 12
2. 藥品及耗材來源 14
三、雌激素代謝產物分析 15
1. 分析使用儀器 15
2. 分析條件設定 15
3. 雌激素代謝物的retention time與被激發電位 17
4. 雌激素代謝產物之標準品配製與標準曲線製作 18
四、肌酸酐濃度測定 (Creatinine assay) 20
1. 吸光值測定 20
2. 濃度計算 21
五、 飼料脂肪酸分析 21
1. 分析使用儀器 21
2. 脂肪酸萃取 21
3. 分析條件設定 22
4. 藥品來源 22
六、統計方法 23
第四章、尿液雌激素代謝產物萃取方法建立-結果 24
1. 加酸加熱處理 24
2. 比較加酸加熱處理與酵素β-glucuronidase/ sulfatase處理 27
第五章、尿液雌激素代謝產物萃取方法建立-討論 28
第六章、尿液雌激素代謝產物分析 32
第一節 生殖期 (Reproductive age) 雌鼠尿液雌激素代謝產物分析 32
一、實驗動物分組 32
二、實驗飼料配製 32
三、分析結果 35
第二節 高n-9與n-6脂肪酸飲食之懷孕期母鼠雌激素代謝產物分析 38
一、實驗動物分組 38
二、實驗飼料配製 38
三、分析結果 41
第三節 高脂與低脂飲食之懷孕期母鼠雌激素代謝產物分析 47
一、實驗動物分組 47
二、實驗飼料配製 48
三、分析結果 50
第七章、尿液雌激素代謝產物分析-討論 60
一、高脂飲食對雌鼠尿液中total estrogen metabolites的影響 60
二、高n-6脂肪酸飲食對16α-OHE1的影響 61
三、高n-6脂肪酸飲食對雌鼠尿液中雌激素 (estrogen) 的影響 64
四、高n-6脂肪酸飲食對2-OHE1的影響 65
五、高脂飲食對hydroxyestrogen甲基化的影響 65
六、不同脂肪酸種類之飲食對雌激素代謝的影響 68
第八章、總結 70
參考文獻. 71

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