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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 黃青真(Ching-Jang Huang) | |
dc.contributor.author | Szu-Chi Liao | en |
dc.contributor.author | 廖思齊 | zh_TW |
dc.date.accessioned | 2021-06-17T01:57:28Z | - |
dc.date.available | 2019-07-27 | |
dc.date.copyright | 2017-07-27 | |
dc.date.issued | 2017 | |
dc.date.submitted | 2017-07-20 | |
dc.identifier.citation | Adams, L. A., Angulo, P., & Lindor, K. D. (2005). Nonalcoholic fatty liver disease. CMAJ, 172(7), 899-905.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/67909 | - |
dc.description.abstract | 近代國人平均壽命逐年增加,然而卻因年輕世代的生育率日漸低迷,使得人口結構愈趨失衡,高齡化社會遂成為我國的重要議題。在許多影響老年人生活品質的因素中,肌少症造成的危害最為直接且嚴重。因此,如何維持年長者的肌肉質量與功能,是科學界急需解決的課題。在眾多可能的治療方法中,雌激素相關受體α(ERRα)被視為最有希望的藥物目標之一。當該受體被活化時,可以提升骨骼肌中的粒線體增殖與活性,進而提升肌肉功能。由於山苦瓜過去被證實能夠提升小鼠骨骼肌中的粒腺體生合成指標基因表現,且其中重要的活性物質葫蘆烷型三萜類與可能是ERRα內源性配體的膽固醇有相似結構,因此本研究的假說為山苦瓜能夠藉由調節ERRα的活性以提升骨骼肌粒線體的增生與功能。
本研究設計了兩階段的實驗以驗證假說。在第一部分中,我們採用了轉錄活化實驗測試山苦瓜的乙酸乙酯萃取物是否能調節ERRα的活性。結果顯示,試驗採用的五種山苦瓜樣品皆能有效降低ERRα的轉錄活性。進一步以三種山苦瓜的活性成分檢視後,顯示共軛亞麻油酸具有最佳的調節效果。於第二部分的實驗,我們將山苦瓜凍乾粉末加入了高脂飼料中,檢視以飲食誘發肥胖的C57BL/6J小鼠骨骼肌中ERRα及其下游基因的表現。最終確認小鼠攝食山苦瓜P81品系後,比目魚肌中的粒腺體動態平衡指標mitofusin 2 mRNA表現顯著提升。 總結以上,本研究結果支持山苦瓜內含成分可以調節雌激素相關受體α活性的假說,並且在肥胖小鼠模式中能提升部分肌肉的mitofusin 2表現,顯示可能具有維持粒線體功能與健康的潛力。 | zh_TW |
dc.description.abstract | The aging of our population has become one of the major concerns in Taiwan. Sarcopenia, the degenerative loss of skeletal muscle mass, quality and strength associated with aging, is one of the most important causes of functional decline and loss of independence in the elderly. Maintaining the mass and functions of skeletal muscles is hence a key issue for healthy aging. Estrogen-related receptor α, ERRα, is required for the activation of mitochondrial genes, and known to regulate genes of mitochondrial biogenesis and energy metabolism. The essential role of ERRα in skeletal muscle fitness and regeneration has been demonstrated. Previously, wild bitter gourds (WBG) were found to elevate mitochondrial biogenesis markers in skeletal muscles in mice. As some active compounds in WBG shows some structural similarity to the endogenous ligand of ERRα, we hypothesize that WBG can promote mitochondrial activity by regulating ERRα. A cell-based ERRα transactivation assay and an animal experiment were conducted to test this hypothesis.
ERRα ligand binding domain was first cloned and then used to develop the cell-based transactivation assay. In this assay, XCT790, a synthetic inverse agonist, activated ERRα at low concentrations, but showed antagonism at higher ones. Genistein, a reported ERRα ligand, dose-dependently inhibited transactivation. A total of five samples, including four cultivars, 1758, P81, 55M and H4, and the product of P81 hydrolyzed at 50℃, P81_50, were tested in this assay. The ethyl acetate extracts of the WBGs also showed dose-dependently inhibitions which were to a greater extent than that of genistein. All the five samples exhibited comparable effects. Among the three active constituents of WBG, α-eleostearic acid, a conjugated linolenic acid showed the highest activity. In the mice study, groups of C57BL/6J mice were fed the high fat diet supplemented with 4% of lyophilized WBG powder for four weeks. The mRNA gene expressions of ERRα and its downstream genes in the skeletal muscles were analyzed. Among these, the mitochondrial dynamics marker, mitofusin 2, which contains ERRα response element within its promoter, was significantly up-regulated in the soleus muscle of mice fed the P81 cultivar of WBG. In conclusion, this study provided preliminary evidence that constituents of WBG might regulate ERRα activity, and promote mitofusin 2 mRNA expression in soleus muscle of high-fat diet fed mice. These results suggest that WBG might have health benefit through the modulation of ERRα. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T01:57:28Z (GMT). No. of bitstreams: 1 ntu-106-R04b22063-1.pdf: 11975052 bytes, checksum: c8c9b4be1e5b9fbed020273d57520829 (MD5) Previous issue date: 2017 | en |
dc.description.tableofcontents | 中文摘要 I
Abstract II 縮寫對照表 IV 總目錄 VI 圖目錄 X 表目錄 XII 第一章 緒論 1 第一節 前言 1 第二節 文獻回顧 2 一、 肌少症 2 二、 骨骼肌 4 2.1 肌纖維 4 2.2 肌肉可塑性(Muscle Plasticity) 5 2.3 肌肉前驅細胞的增生與分化(Myoblast Proliferation and Differentiation) 5 三、 雌激素相關受體α (Estrogen-Related Receptor α, ERRα) 7 3.1 雌激素相關受體的異形體(Isoforms of ERR) 8 3.2 ERRα的配體(Ligands) 9 3.3 ERRα與PGC-1α 11 3.4 ERRα與粒線體 12 3.5 ERRα與骨骼肌 12 3.6 ERRα被抑制或過度活化時的動物表現型(phenotype) 13 四、 PGC-1α (Peroxisome proliferator-activated receptor gamma coactivator 1-α) 15 4.1 老化對於PGC-1α的影響及其可能的機制 15 4.2 PGC-1α與肌肉蛋白質降解、細胞自噬(Proteolysis and Autophagy) 16 4.3 PGC-1α與粒線體的動態平衡(Mitochondrial Dynamics) 18 五、 粒線體 18 5.1 粒線體生合成(Mitochondrial Biogenesis) 18 5.2 粒線體動態(Mitochondrial Dynamics) 19 5.3 粒線體與肌少症 21 六、 山苦瓜 22 6.1 葫蘆烷型三萜類分子(Cucurbitane-type Triterpenoids)和山苦瓜中其他活性成分 22 6.2 山苦瓜對骨骼肌的功效 24 第三節 研究假說與實驗架構 26 一、 研究假說 26 二、 實驗架構 27 第二章 雌激素相關受體α轉錄活化活性分析 28 第一節 前言 28 第二節 材料與方法 30 一、 試劑與器材 30 雌激素相關受體α 基因選殖與質體製備 30 轉錄活化活性分析 31 二、 實驗方法 35 雌激素相關受體α基因選殖與質體建構 35 (一) 聚合酶連鎖反應(Polymerase chain reaction, PCR) 36 (二) DNA電泳 37 (三) Gel/PCR DNA純化 37 (四) 重組DNA — T&A Cloning 38 (五) 重組DNA — ERRα與pBK-CMV GAL4 Vector 38 (六) 轉形作用(Transformation)與單株菌落篩選(Single colony selection) 39 (七) 小量質體抽取 39 (八) 中量質體抽取 40 轉錄活化活性分析 40 (一) 新鮮山苦瓜處理 41 (二) 山苦瓜內源性酵素水解 41 (三) 山苦瓜乙酸乙酯萃取 41 (四) CHO-K1細胞培養 41 (五) 短暫轉染 42 (六) 以MβCD降低細胞膽固醇含量 43 (七) 細胞存活率分析—MTT染色法 43 (八) 統計分析 43 第三節 結果 45 雌激素相關受體α 基因選殖及質體製備 45 (一) PCR產物確認 45 (二) Colony PCR 45 (三) 質體DNA定序與限制酶圖譜 45 轉錄活化活性分析 46 (一) PGC-1α之活化效果 46 (二) 以MβCD降低細胞膽固醇含量對ERRα活性的影響 46 (三) 外加膽固醇之效應 47 (四) 標準品Genistein及XCT790之效應 47 (五) 山苦瓜乙酸乙酯萃取物的影響 48 (六) 山苦瓜中活性成分的影響 49 第四節 討論 61 一、 ERRα基因選殖 61 二、 MβCD處理及細胞膽固醇濃度對ERRα轉錄活性之影響 61 三、 標準品金雀異黃酮及XCT790 64 四、 山苦瓜乙酸乙酯萃取物及成分物質調節ERRα活性的潛力 66 五、 與Docking結果的比較 69 第五節 結論 72 第三章 山苦瓜添加對攝食高脂飼料小鼠的影響 73 第一節 前言 73 第二節 實驗設計 74 第三節 材料與方法 75 一、 動物飼養 75 二、 飼料配製 75 2.1 Chow Diet 75 2.2 高脂飼料 75 2.3 苦瓜飼料 76 三、 禁食血清樣本蒐集 79 四、 動物犧牲與取樣 79 五、 血清葡萄糖分析 79 六、 血清三酸甘油酯分析 80 七、 血清膽固醇分析 80 八、 血清胰島素分析 80 九、 胰島素阻抗指標計算 81 十、 肌肉組織mRNA表現分析 81 10.1 總mRNA抽取 81 10.2 總mRNA反轉錄為cDNA 81 10.3 Quantitative Real-time PCR 82 十一、 統計分析 83 第四節 結果 85 一、 動物表徵 85 (一) 生長曲線與體重增加量 85 (二) 攝食量、能量攝取與能量效率 85 二、 器官與組織重量 86 (一) 絕對重量 86 (二) 相對重量 86 三、 血清生化分析 87 (一) 血清葡萄糖分析 87 (二) 血清三酸甘油脂分析 87 (三) 血清膽固醇分析 88 (四) 血清胰島素與胰島素阻抗指標 89 四、 粒線體相關基因及ERRα下游基因表現 89 (一) 腓腸肌基因表現 90 (二) 比目魚肌基因表現 91 第五節 討論 107 一、 動物體重與脂質堆積 107 二、 胰島素阻抗與肝臟脂質累積 108 三、 骨骼肌組織中mRNA表現 109 第六節 結論 115 第四章 綜合討論與總結論 116 第一節 綜合討論 116 第二節 總結論 120 第五章 參考文獻 121 附錄一 電腦模擬預測山苦瓜活化ERRα之潛力 136 附錄二 水解前後山苦瓜萃取物中三萜類含量 141 附錄三 水解前後山苦瓜萃取物中CLN含量 142 附錄四 山苦瓜乙酸乙酯萃物活化PPARδ轉錄活性測試 143 | |
dc.language.iso | zh-TW | |
dc.title | 山苦瓜作為雌激素相關受體α調節劑調控小鼠骨骼肌粒線體之活性 | zh_TW |
dc.title | Wild Bitter Gourd Regulates Mitochondrial Activity in the Skeletal Muscle of Mice by Functioning as an Estrogen-Related Receptor α Modulator | en |
dc.type | Thesis | |
dc.date.schoolyear | 105-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 呂紹俊,蘇慧敏,張美鈴,林甫容 | |
dc.subject.keyword | 肌少症,雌激素相關受體α,粒線體,山苦瓜,共軛亞麻油酸, | zh_TW |
dc.subject.keyword | sarcopenia,estrogen-related receptor α,mitochondria,wild bitter gourd,conjugated linolenic acid, | en |
dc.relation.page | 143 | |
dc.identifier.doi | 10.6342/NTU201701751 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2017-07-21 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科技學系 | zh_TW |
顯示於系所單位: | 生化科技學系 |
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