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研究生:顏于婷
研究生(外文):Yu-Ting Yen
論文名稱:在體外及體內模式下探討薏苡殼及種皮之乙醇萃取物對子宮肌瘤細胞的影響
論文名稱(外文):The effects of adlay hull and testa ethanolic extracts on the uterine leiomyoma cells in vitro and in vivo
指導教授:江文章江文章引用關係
指導教授(外文):Wenchang Chiang
口試委員:施純光吳啟豪
口試日期:2015-07-30
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:食品科技研究所
學門:農業科學學門
學類:食品科學類
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:111
中文關鍵詞:子宮肌瘤性激素誘導子宮肌瘤小鼠模式薏苡豆固醇
外文關鍵詞:uterine leiomyomahormone-induced uterine fibroid mouse modeladlaystigmasterol
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子宮肌瘤是女性最常發生的良性腫瘤,臨床上約有20%的女性患有子宮肌瘤,少數患者會有劇烈腹痛、異常出血及不孕等症狀。本研究室先前研究顯示薏苡籽實萃取物具有抑制腫瘤及惡性肉瘤細胞生長之功效,亦有研究證實薏苡殼甲醇萃取物可藉由抑制芳香酶的活性以調節雌激素及黃體素的生成量。本研究的目的為:(一) 以細胞模式探討薏苡殼與薏苡種皮乙醇萃取物的不同次區分層對於子宮肌瘤的生長抑制效果;(二)以動物模式探討薏苡殼與薏苡種皮乙醇萃取物對於性激素誘導子宮肌瘤小鼠模式內分泌調節及抗發炎反應之效果。結果顯示薏苡殼與薏苡種皮乙醇萃取物乙酸乙酯區分層對於子宮肌瘤細胞均有抑制效果,而其中以薏苡殼乙醇萃取物沉澱層之乙酸乙酯次區分層(AHE-S-ea)對於子宮肌瘤細胞株的生長抑制效果最為顯著,培養2天及4天後細胞存活率分別為67.3±0.6%及53.3±3.7%;AHE-S-ea對於子宮肌瘤初代細胞也有顯著的抑制效果,培養2天及4天後細胞存活率為51.0±9.5%及34.8±8.5%。動物實驗的結果指出薏苡殼乙醇萃取液乙酸乙酯次區分層(AHE-ea)及豆固醇(Stigmasterol)可有效降低子宮肌瘤動物的子宮臟器係數,也可降低動物體內雌激素與黃體酮含量,發炎因子的測定結果顯示,AHE-ea可有效降低子宮肌瘤動物血清中TNF-α的含量,因此AHE-ea可能也透過免疫方面的調節來抑制子宮肌瘤。未來可繼續探討薏苡殼乙醇萃取物之次區分層對於預防或輔助治療子宮肌瘤之作用機制,以提供更多女性保健食品開發之科學證據。

Uterine leiomyomas are the most common benign tumors in women, with up to 20% of this population being clinically apparent. Patients with uterine leiomyomas often experience abnormal uterine bleeding, pelvic pain or pressure, infertility and recurrent pregnancy loss. Previous studies have shown adlay extract possesses anti-tumor activity and inhibit sarcoma cell proliferation and tumor progression. Many studies also confirmed that the methanol extracts of Adlay hull regulate the amount of estrogen and progesterone generate by inhibiting the activity of aromatase. Therefore, the purpose of this study are (1) to investigate the inhibition effects and mechanisms of the adlay hull and testa ethanolic extracts on fibroid growth and extracellular matrix accumulation in vitro. (2) to confirm the effects of the adlay ethanolic extracts on hormone-induced uterine fibroid mouse model for endocrine regulation and the anti-inflammatory effect in vivo. The results showed that adlay hull and testa ethanolic extracts ethyl acetate subfraction have inhibitory effect on leiomyoma cells and the ethyl acetate fraction of sediment of adlay hull ethanolic extracts (AHE-S-ea ) has the most significant inhibitory effect on leiomyoma cells growth. The results of animal experiments indicated the ethyl acetate fraction of adlay hull ethanolic extracts (AHE-ea) and stigmasterol can effectively reduce the uterine coefficient in uterine fibroid animals, they can also reduce the serum content of estrogen and progesterone. The imflammatory factor experiment showed that AHE-ea can improve TNF-α in mouse with uterine fibroids , thus AHE-ea may inhibit uterine fibroids through regulating immunity. In conclusion, AHE-ea and stigmasterol have the potential for regulating the uterine leiomyoma models in vitro and in vivo.

目 錄


摘要 ………………………………………………………………………………… II
Abstract ……………………………………………………………………………… III
縮寫對照表………………………………………………………………………… IV
目錄………………………………………………………………………………… VI
圖目錄……………………………………………………………………………… IX
表目錄……………………………………………………………………………… XII

第一章、前言……………………………………………………………………… 1

第二章、文獻回顧………………………………………………………………… 2
一、子宮肌瘤(Uterine Leiomyoma)
(一)子宮生理結構……………………………………………………… 2
(二)流行病學…………………………………………………………… 2
(三)病因學……………………………………………………………… 4
(四)臨床處置…………………………………………………………… 6
(五)子宮肌瘤輔助療法之研究………………………………………… 6
二、薏苡籽實介紹及相關研究
(一)薏苡簡介…………………………………………………………… 9
(二)薏苡之生理機能性研究…………………………………………… 10
(三)薏苡籽實及其萃取物或區分層對固醇類荷爾蒙調控之研究…… 16
(四)薏苡籽實中的化合物對抗腫瘤影響之相關研究………………… 21

第三章、研究動機與實驗架構…………………………………………………… 35
一、研究動機………………………………………………………………… 35
二、實驗架構………………………………………………………………… 36


第四章、以體外模式篩選薏苡籽實萃取物抑制子宮肌瘤細胞生長及細胞外基質累積之活性次區分層以及其活性成分分析……………………………………… 39
一、材料與方法
(一)薏苡殼、種皮乙醇萃取物之製備……………………………… 39
(二)薏苡殼、種皮乙醇萃取物之植物固醇分析…………………… 42
(三)細胞培養………………………………………………………… 42
(四)細胞數之計算…………………………………………………… 43
(五)細胞存活率分析─MTT assay…………………………………… 43
(六)細胞型態分析─結晶紫染色……………………………………… 46
(七)統計分析………………………………………………………… 46
二、結果
(一)薏苡殼、種皮乙醇萃取物及不同活性區分層之製備……………47
(二)薏苡殼、種皮乙醇萃取物及不同活性區分層之植物固醇分析…50
(三)薏苡殼乙醇萃取物之次區分層對子宮肌瘤細胞生長抑制之相關研
究…………………………………………………………………… 50
(四)薏苡種皮乙醇萃取物之次區分層對子宮肌瘤細胞生長抑制之相關
研究………………………………………………………………… 61
(五)薏苡殼及薏苡種皮乙醇萃取物乙酸乙酯區分層對子宮肌瘤及正常
子宮平滑肌細胞株生長型態之影響……………………………… 71
(六)雌性類固醇激素對子宮平滑肌細胞促進增生之相關研究……… 74

第五章、以體內模式探討薏苡殼乙醇萃取物乙酸乙酯層對於子宮肌瘤小鼠性激素表現及發炎反應的影響…………………………………………………………… 77
一、材料與方法
(一)性激素誘導子宮肌瘤動物模式…………………………………… 77
(二)實驗動物及分組…………………………………………………… 77
(三)組織樣本收集與分析方法…………………………………………78
(四)統計分析…………………………………………………………… 81
二、結果
(一)薏苡殼萃取物乙酸乙酯層對動物生長之影響…………………… 81
(二)薏苡殼萃取物乙酸乙酯區分層對性荷爾蒙分泌之影響………… 86
(三)薏苡殼萃取物乙酸乙酯區分層對動物發炎反應之影響………… 89

第六章、綜合討論………………………………………………………………… 91

第七章、結論……………………………………………………………………… 98

參考文獻…………………………………………………………………………… 99


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