薏苡殼萃取物在體外及體內試驗對於雄烯二酮分泌的影響

Ya-Ting Wu; 吳雅婷

請用此 Handle URI 來引用此文件： http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/22831

完整後設資料紀錄

DC 欄位	值	語言
dc.contributor.advisor	江文章
dc.contributor.author	Ya-Ting Wu	en
dc.contributor.author	吳雅婷	zh_TW
dc.date.accessioned	2021-06-08T04:29:54Z	-
dc.date.copyright	2011-08-22
dc.date.issued	2011
dc.date.submitted	2011-08-17
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dc.identifier.uri	http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/22831	-
dc.description.abstract	雄烯二酮(androstenedione, AD)主要在腎上腺和卵巢生成，是女性合成睪固酮與雌激素的原料，若分泌不平衡便會產生失調。多囊性卵巢症候群(polycystic ovary syndrome, PCOS)是育齡婦女中常見的內分泌疾病，診斷標準為以下三項中至少有兩項：(1)寡排卵或不排卵、(2)臨床或生化指標有高雄性激素的表現與(3)卵巢呈現多囊性狀；同時因為持續的高雄性激素表現使得發生胰島素阻抗、第二型糖尿病、子宮內膜癌的機率增加。文獻指出薏苡(Coix lachryma-jobi L. var. ma-yuen Stapf)殼(adlay hull, AH)甲醇萃取物(AH methanolic extract, AHM)可抑制雌性大鼠雌二醇的分泌，且降低雄性大鼠的睪固酮生成。本研究以人類絨毛膜促性腺激素(human chorionic gonadotropin, hCG)誘發卵巢初代包膜細胞(theca cell, TC)過量分泌雄烯二酮模式評估薏苡籽實萃取物之抑制活性，進而以動物模式驗證，並篩選有效次區分物與有效化合物。結果顯示，薏苡殼乙醇萃取物(AH ethanolic extract, AHE)具顯著抑制效果，其中以乙酸乙酯區分層(ethyl acetate fraction of AHE, AHE-EA)活性最佳，且可降低PCOS模式動物血清中雄烯二酮含量、禁食血糖與血清胰島素，同時改善卵巢中的氧化壓力；而其次區分物中以AHE-EA-H抑制活性最佳。利用液相層析串聯質譜(liquid chromatography-mass spectrum-mass spectrum, LC-MS-MS)比對此層所得物質進行確效，證實化合物3號、4號、5號、6號和9號在hCG的刺激下具有抑制雄烯二酮的效果；這五個化合物皆可視為有效成分，其中化合物4號在100 μM處理下可抑制82.7%的雄烯二酮分泌，可做為薏苡籽實中調節雄烯二酮分泌的指標物質。	zh_TW
dc.description.abstract	Androstenedione (AD) is mainly synthesized in adrenal glands and ovaries, which is precursors of testosterone and estrogen in women, while imbalance secretion of AD leads to dysfunction of endocrine system. Polycystic ovary syndrome (PCOS) commonly occurs in childbearing women, and the diagnostic criteria include at least 2 subjects with following 3 events: (1) oligo-ovulation or anovulation; (2) hyperandrogenism clinically or in biomarkers; and (3) polycystic ovaries. Preceding high expression of androgens increase the risks of insulin resistance, type 2 diabetes, and endometrial cancer. Adlay (Coix lachryma-jobi L. var. ma-yuen Stapf) hull (AH) methanolic extract (AHM) was reported to suppress estradiol release from female rats and decrease level of testosterone in male ones. In the present investigation, the inhibitory effects of extracts from adlay seeds on AD secreted from human chorionic gonadotropin (hCG)-stimulated primary rat theca cell (TC). Further, the effective extracts in vitro were verified in vivo, and the active sub-fractions and effective components were elucidated. Results demonstrated that AH ethanolic extract (AHE) possessed significantly inhibitory capacity, and ethyl acetate fraction from AHE (AHE-EA) showed superior activity, which lowered the levels of serum AD, fasting blood glucose, and serum insulin; meanwhile the oxidative stress in ovaries was improved in PCOS animal model. H-sub-fraction from AHE-EA showed most favorable inhibitory potential in vitro, and was analyzed by liquid chromatography (LC)-mass spectrum (MS)-MS. Compounds 3, 4, 5, 6, and 9 significantly suppressed AD release in vitro among them, 4 inhibited 82.7% of AD at 100 μM, and may be regarded as indicator in adlay seeds on modulation of AD secretion.	en
dc.description.provenance	Made available in DSpace on 2021-06-08T04:29:54Z (GMT). No. of bitstreams: 1 ntu-100-R98641033-1.pdf: 2504527 bytes, checksum: 7db8c12451411c85d3e6da6825074502 (MD5) Previous issue date: 2011	en
dc.description.tableofcontents	目錄圖目錄 XII 表目錄 XV 第一章、前言 1 第二章、文獻回顧 2 一、女性內分泌生殖系統介紹 2 1.卵巢(Ovary) 2 (1) 包膜細胞 3 2.卵巢類固醇性激素合成路徑(Steroidogenesis) 4 3.兩個細胞，兩個性促激素學說(The two-cell, two-gonadotrophin theory) 5 4.月經週期 6 二、多囊性卵巢症候群(Polycystic ovary syndrome, PCOS) 7 1.起源與歷史 7 2.診斷標準 7 (1) 1990年的診斷標準(The 1990 National Institutes of Health Criteria) 7 (2) 2003年的診斷標準(The 2003 ESHRE/ASRM Criteria) 8 (3) 2006年的診斷標準(The AES 2006 Criteria) 8 (4) 應排除的相關疾病 8 3.臨床表徵 9 (1) 臨床上或生化上血中雄性素過高 ( Hyperandrogenism) 9 (2) 胰島素阻抗(Insulin resistance) 9 (3) 發炎(Inflammation) 10 4. 臨床處理 10 (1) 藥物治療 10 (2) 非藥物治療 13 三、薏苡籽實介紹與相關研究 17 1.薏苡簡介 17 2.薏苡之生理機能性研究 18 3.薏苡籽實及其萃取物對類固醇激素分泌之研究 22 4.薏苡籽實及其萃取物對子宮收縮以及婦癌腫瘤之研究 24 5.自薏苡殼中分離出來的植物化學成分 25 第三章、研究動機、目的與架構 27 一、研究動機 27 二、研究目的 27 三、研究架構 27 第四章、以體外初代細胞模式篩選薏苡籽實萃取物抑制雄烯二酮活性以及其活性成分分析 31 一、材料與方法: 31 1.薏苡籽實各部位乙醇萃取物及不同極性區分物的製備 31 (1) 薏苡殼乙醇萃取物及不同極性區分物的製備 31 (2) 薏苡種皮乙醇萃取物的製備 32 (3) 薏仁麩皮乙醇萃取物的製備 32 (4) 精白薏仁乙醇萃取物的製備 32 二、大鼠卵巢包膜細胞培養 32 1.實驗動物 32 2.卵巢包膜間隙細胞的製備 32 3.細胞數及存活率之計算 33 4.免疫螢光染色 36 5.激素之測定(雄烯二酮) 36 (1) 激素萃取 36 (2) 進行EIA kit測定 36 三、薏苡殼活性成分分析 37 1.薄層分析法 37 2.總酚含量測定 38 3.總類黃酮含量測定 38 4.高效能液相層析儀串聯質譜儀分析 39 5.統計分析 39 四、結果 40 1.以免疫螢光法觀察大鼠卵巢包膜細胞中的Vimentin及DAPI的表現 40 2.人類絨毛膜性促素對卵巢包膜細胞刺激分泌雄烯二酮之時程效應 40 3.人類絨毛膜性促素對卵巢包膜細胞刺激分泌雄烯二酮之劑量效應 40 4.薏苡籽實乙醇萃取物及其區分物對卵巢包膜細胞分泌雄烯二酮之效應 41 (1) 薏苡籽實不同部位乙醇萃取物對卵巢包膜細胞分泌雄烯二酮之效應 41 ( 2) 薏苡殼乙醇萃取物之不同極性區分物對卵巢包膜細胞分泌雄烯二酮之效應 41 (3) 不同劑量的薏苡殼乙醇萃取物乙酸乙酯層對卵巢包膜細胞分泌雄烯二酮之效應 41 (4) 薏苡殼乙醇萃取物乙酸乙酯層之次區分物對卵巢包膜細胞分泌雄烯二酮之效應 42 (5) 薏苡殼乙醇萃取物乙酸乙酯層B、H、I、J、L、N次區分層在adenylyl cyclase 刺激劑-forskolin、 cAMP 類似物-8-Br-cAMP 活化cAMP-PKA、親水性膽固醇25-OH-cholesterol可直接進入粒線體內路徑下對大鼠卵巢包膜細胞分泌孕酮之效應 42 5.薏苡殼乙醇萃取物活性區分物之成分分析 43 (1) 薏苡殼乙醇萃取物乙酸乙酯區分物及其次區分物總酚及總類黃酮含量 43 (2) 經高效能液相層析串聯質譜儀分析薏苡殼乙醇萃取物之乙酸乙酯層之其次區分物中酚類化合物之定性結果 43 6.薏苡殼中所含類黃酮及其它化合物對卵巢包膜細胞分泌雄烯二酮之效應 43 第五章、體內試驗進行驗證薏苡殼萃取物乙酸乙酯區分層對於多囊性卵巢症候群雄烯二酮以及其相關症狀的影響 54 一、材料與方法 54 1.脫氫表固酮誘導多囊性卵巢症候群動物模式 54 2.實驗動物 54 3.動物分組 54 4.動情週期判定 54 5.組織樣本收集與分析方法 55 (1) 禁食血糖含量測定 55 (2) 血液分析 56 (3) 卵巢分析 59 6.統計分析 61 二、結果 62 1.薏苡殼萃取物乙酸乙酯區分層對動物生長、攝食量以及組織臟器之影響 62 2.薏苡殼萃取物乙酸乙酯區分層對卵巢組織及血液相關生化值之影響 62 (1) 血清肝臟酵素GOT、GPT含量 62 (2) 血清雄烯二酮、雌二醇以及助孕酮含量 62 (3) 禁食血糖變化、血清胰島素含量 63 (4) 卵巢脂質過氧化物含量 63 (5) 血清超氧歧化酶及過氧化氫酶 63 (6) 血清C反應蛋白 63 3.卵巢切片之組織學觀察 64 第六章、綜合討論 82 一、薏苡籽實乙醇萃取物對人絨毛膜性促素(hCG)誘導卵巢包膜細胞過度分泌雄烯二酮之探討 82 二、薏苡殼乙醇萃取物乙酸乙酯區分層對以脫氫睪固酮誘導多囊性卵巢症候群動物模式之探討 83 第七章、結論 89 參考文獻 90
dc.language.iso	zh-TW
dc.subject	包膜細胞	zh_TW
dc.subject	雄烯二酮	zh_TW
dc.subject	多囊性卵巢症候群	zh_TW
dc.subject	薏苡殼	zh_TW
dc.subject	adlay hull	en
dc.subject	theca cell	en
dc.subject	polycystic ovary syndrome	en
dc.subject	androstenedione	en
dc.title	薏苡殼萃取物在體外及體內試驗對於雄烯二酮分泌的影響	zh_TW
dc.title	Effect of Adlay Hull Extracts on the Secretion of Androstenedione In Vitro and In Vivo	en
dc.type	Thesis
dc.date.schoolyear	99-2
dc.description.degree	碩士
dc.contributor.coadvisor	夏詩閔
dc.contributor.oralexamcommittee	吳兩新,張素瓊,陳美州
dc.subject.keyword	雄烯二酮,多囊性卵巢症候群,薏苡殼,包膜細胞,	zh_TW
dc.subject.keyword	androstenedione,polycystic ovary syndrome,adlay hull,theca cell,	en
dc.relation.page	105
dc.rights.note	未授權
dc.date.accepted	2011-08-17
dc.contributor.author-college	生物資源暨農學院	zh_TW
dc.contributor.author-dept	食品科技研究所	zh_TW
顯示於系所單位：	食品科技研究所

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