臺灣博碩士論文加值系統

流行病學研究指出膳食中單碳葉酸及膽鹼營養素攝取不足可能會影響附基因體的甲基化修飾。懷孕婦女單碳膽素及葉酸需求量大，不僅需維持母體單碳所需，更是需要提供胎兒足夠的單碳代謝環境以利胎兒生長發育，而有癌症風險因子之孕婦其單碳營養素攝取結合單碳代謝酵素基因多型性變異(MTHFR C677T)對孕期發炎反應及印記基因甲基化的相關性仍尚未清楚。本研究目標：有親代癌症風險的孕婦其孕期單碳葉酸及膽素營養狀態與發炎反應及印記基因甲基化的相關性探討。由台大醫院及樂寶兒診所聯合招募50位懷孕初期的受試者，藉由填寫飲食頻率問卷及24小時飲食回憶記錄，了解其飲食單碳營養素攝取狀況，同時收取血液檢體測量其血漿單碳營養素與發炎反應指標並萃取淋巴球DNA檢測MTHFR C677T基因型與印記基因IGF2/H19和總基因體(LINE-1)的甲基化。結果顯示，孕婦膽素攝取不足比例為52% (平均攝取量：434±228 mg/day；足夠攝取建議值: 410 mg/day)，葉酸缺乏率為16% (平均攝取量：1157±946 ug/day；每日飲食建議攝取量: 600 ug/day)，Vitamin B6缺乏率為64% (平均攝取量：6.4±15.3 mg/day；每日飲食建議攝取量: 1.9 mg/day)。家族史肥胖的孕婦相較於正常孕婦組，有顯著高的總膽素(P = 0.048)及葉酸(P = 0.002)攝取，但其血漿膽素濃度顯著較低(P = 0.049)。MTHFR CT/TT基因型組孕婦其飲食總膽素(P = 0.047)、Phosphotidylcholine (P = 0.020)、Sphingomyelin (P = 0.022) 攝取顯著高於CC基因型組，但血漿葉酸(P = 0.049)及甜菜鹼(P = 0.023)濃度卻顯著較低。而發炎反應中，血漿Vitamin B12(r = -.333, P < 0.001)及甜菜鹼(r = -.227, P = 0.013)濃度與CRP呈顯著負相關。對於DNA甲基化的影響，飲食葉酸(r = -.327, P = 0.028)、甜菜鹼(r = -.315, P = 0.035)攝取及血漿甜菜鹼濃度(r = -.200, P = 0.033)皆與總基因體 LINE-1 甲基化呈顯著負相關。但血漿Vitamin B12(r = .203, P = 0.034)及甜菜鹼(r = .213, P = 0.023)濃度與印記基因 IGF2/H19甲基化呈顯著正相關，相關原因目前尚未清楚，仍需要更多研究進行探討。

Dietary intake of folate and choline appear to be inversely related to epigenetic process in epidemiological studies. Pregnant women one-carbon choline and folate requirements, not only needed to maintain maternal one carbon metabolism, but also need to provide enough of one-carbon nutritional environment for fetal to growth and development. Folate and choline of status in pregnant with parental cancer diseases risks and combined with one carbon metabolism enzyme gene polymorphism mutation (MTHFR C677T) correlation of inflammation and imprinted gene methylation in pregnancy has not been clearly. Aims of the study were to investigate the associations of pregnancy one carbon nutrients status with cancer risk factors, inflammatory response and methylation of imprinted gene. Fifty pregnant women were recruited from multiple clinic centers and NTU hospital to participate through the study. Blood samples from the pregnant women were collected for measurements of plasma one-carbon nutritients and inflammation levels. Lymphocytic DNAs were extracted for methylenetetrahydrofolate reductase (MTHFR) genotyping and for methylation status of imprinted gene IGF2/H19 (insulin-like growth factor-2) and LINE-1 (long interspersed nucleotide elements). Dietary data were collected by FFQ and analyzed for nutrient intakes. The data revealed that deficiency rates of total choline, folate and vitamin B6 intake (< RDA/AI) among the first trimester pregnant women were 52%, 16% and 64%, respectively. Historical parental obesity in relative to normal-weight parents was associated with higher intake levels of folate and total choline, and with significantly lower plasma free choline levels in pregnant offsprings. MTHFR 677CT/TT genotypes in relative to CC wildtype were associated with higher intakes of phosphotidylcholine and total choline, and with lower plasma folate and betaine levels. Inflammation results show that plasma vitamin B12 and betaine concentrations were negatively correlated with CRP levels. Dietary intake of folate and betaine were a significant negative correlation with LINE-1 methylation, but plasma vitamin B12 and betaine concentrations were positively correlated with the methylation of imprinted gene IGF2/H19. Detailed mechanism is not clear, still needs more research to explore the related causes.

摘要
Abstrate
誌謝
表目錄
縮寫表 (Abbreviation)
第一章 前言
第二章 文獻回顧
一、 單碳營養素之營養生化代謝作用
(一) 單碳代謝循環
(二) 單碳營養素與基因調控
(三) 基因型及DNA甲基化與癌症之風險性
二、 惡性腫瘤發展相關分子機制
(一) Insulin-like growth factor-2 (IGF2) 訊息傳遞分子機制
三、 附基因體甲基化修飾與腫瘤進展
(一) 印記基因- IGF2/H19
(二) 總基因體- LINE-1
四、 孕婦單碳膽鹼營養狀態與癌症發展之相關性
(一) 孕婦單碳代謝酵素基因型對膽鹼營養狀態之影響
(二) 孕婦膽鹼營養狀態對印記基因甲基化修飾及發炎因子之影響
(三) 孕婦單碳營養狀態對癌症之風險性
五、 國人懷孕婦女單碳葉酸、膽鹼營養狀態及需求量
六、 研究目標與假說
第三章 實驗架構與材料方法
一、 實驗設計
二、 實驗材料
(一) 受試者
(二) 一般問卷及飲食問卷
(三) 血液樣本
(四) 儀器設備
(五) 實驗用試藥
三、 實驗方法
(一) 一般問卷與飲食問卷調查
(二) 血樣收集與單核淋巴球分離
(三) 血液單碳營養素測定
(四) 單碳代謝酵素MTHFR C677T 基因型檢測
(五) 總基因體及印記指標之甲基化分析
(六) CRP酵素免疫分析法 (DuoSet ELISA)
四、 統計分析
第四章 實驗結果
一、 受試者基本資料及癌症風險因子之探討
(一) 受試者之癌症高風險因子
(二) 孕婦對於單碳營養素補充劑之使用情形
二、 孕婦每日單碳營養素攝取情形
(一) 每日飲食攝取情形
(二) 診斷單碳營養素攝取情形之指標
(三) 單碳營養素攝取狀況之評估
三、 孕婦第一孕期之血液生化值
(一) 血漿單碳營養狀態
(二) 血漿發炎指標之表現
(三) DNA甲基化狀態
(四) 受試者之MTHFR C677T基因型
四、 孕婦的癌症風險因子與單碳營養狀態之相關性探討
五、 孕婦的癌症風險因子、單碳營養狀態對DNA甲基化及發炎反應之影響
(一) 孕婦的癌症風險因子、單碳營養狀態及發炎反應對LINE-1甲基化的影響
(二) 孕婦的癌症風險因子、單碳營養狀態及發炎反應對印記基因IGF2/H19甲基化的影響
(三) 單碳營養狀態與DNA甲基化之相關性
(四) 孕婦的癌症風險因子、單碳營養狀態與發炎反應之相關性
六、 孕期間的單碳營養狀況及血液生化值之變化
(一) 孕期間飲食單碳營養素攝取變化
(二) 孕期間的血漿單碳營養素、發炎反應及DNA甲基化的變化
第五章 討論
一、 孕婦單碳營養素攝取情形
(一) 診斷單碳營養素攝取情形之指標
(二) 單碳營養素攝取情形
二、 孕婦第一孕期之血液生化值
(一) 血漿單碳營養狀態
(二) 血漿發炎指標之表現
(三) DNA甲基化狀態
(四) MTHFR C677T基因型
三、 孕婦的癌症風險因子與單碳營養狀態之相關性探討
四、 孕婦的癌症風險因子、單碳營養狀態及發炎反應對DNA甲基化之影響
(一) 孕婦的癌症風險因子、單碳營養狀態及發炎反應對LINE-1甲基化的影響
(二) 孕婦的癌症風險因子、單碳營養狀態及發炎反應對印記基因IGF2/H19甲基化的影響
(三) 單碳營養狀態與DNA甲基化之相關性
(四) 孕婦的癌症風險因子、單碳營養狀態與發炎反應之相關性
五、 孕期間的單碳營養狀況及血液生化值之變化
(一) 孕期間飲食單碳營養素攝取變化
(二) 孕期間的血漿單碳營養素、發炎反應及DNA甲基化的變化
第六章 總結
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