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研究生:楊雅婷
論文名稱:紫錐菊萃取物和咖啡酸在斑馬魚脂肪肝形成時的影響
論文名稱(外文):Dry extract of Echinacea purpurea and caffeic acid effect the fatty liver formation in zebrafich
指導教授:陳泓志陳泓志引用關係吳金洌吳金洌引用關係
學位類別:碩士
校院名稱:弘光科技大學
系所名稱:生物科技研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:62
中文關鍵詞:斑馬魚牛油脂肪肝steotosis肝癌中草藥紫錐菊咖啡酸
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台灣民眾得到脂肪肝是一件非常常見的病症。許多人在不知不覺因為不正常飲食及藥物濫用輕易得到脂肪肝。此研究以紫錐菊(Echinacea purpura),一種具有抗癌、抗發炎、抗氧化和可調節免疫的中草藥,試圖抑制已得脂肪肝的成魚斑馬魚。首先以牛油誘導斑馬魚得到脂肪肝,比例分成三種,包括0%、8%、13%,持續餵食120隻成魚斑馬魚一個月;一個月後開始用紫錐菊和咖啡酸處理以得脂肪肝的成魚斑馬魚。總數99隻的成魚斑馬魚分成11組:0%牛油、0%牛油和2.5% 紫錐菊、餵食0%牛油和5% 紫錐菊、0%牛油和0.075mg/ml咖啡酸、0%牛油和0.15mg/ml咖啡酸、8%牛油、8%牛油和2.5% 紫錐菊、8%牛油和5% 紫錐菊、8%牛油和0.075mg/ml咖啡酸、8%牛油和0.15mg/ml咖啡酸、13%牛油;牛油和紫錐菊每天餵食兩次,咖啡酸溶液每天換一次。另外以轉基因斑馬魚(有脂肪肝病徵的魚)餵食紫錐菊和咖啡酸:對照組、2.5% 紫錐菊、5% 紫錐菊、0.075mg/ml咖啡酸、0.15mg/ml咖啡酸。本實驗餵食牛油於斑馬魚後,在基因方面,結果顯示牛油餵食一個月後adiponectin、Apoa1及TNF等三個基因有明顯上升現象。冷凍切片結果顯示8%的牛油餵食可使斑馬魚肝臟脂肪細胞變多,但在13%的牛油餵食後,卻沒有同樣的情形。所以實驗動物以8%的牛油來當作誘導。經過餵食紫錐菊和咖啡酸15天處理已得脂肪肝的斑馬魚,冷凍切片部分發現除了餵食0.15mg/ml咖啡酸的Oil red O的表現有增多情形,其餘紅色油滴皆有減少情形。在基因方面adiponectin、Apoa1及TNF等三個表現量皆因咖啡酸和紫錐菊的原因而下降。
The fatty liver disease is a popular disease in Taiwan. Many people eat by an abnormal dietary or misuse medicine and get the fatty liver (steatosis) unknowingly and easily. This study tries to address the common Chinese herbal Echinacea purpura content-caffeic acid to reduce the adult zebrafish which get fatty liver symptom already. Echinacea purpura is an anticancer, anti-inflammation, anti-oxidation and immunomodulatory properties; the same effects in its content- caffeic acid. First, we must induce fatty liver symptom in adult zebrafish by butter (Saturated fatty acid) high-fat diets. The percentage of butter had divided into three parts, include 0%, 8%, 13%, and feed for one month and more; start to treat the caffeic acid and Echinacea purpura into the different butter treatment of zebrafish after one month. The amounts of 99 zebrafishs were divided into 11 parts: 0% butter control group, 0% butter plus 2.5% Echinacea purpura treatment group, 0% butter plus 5% Echinacea purpura treatment group, 0% butter plus 0.075mg/ml caffeic acid treatment group and 0% butter plus 0.15mg/ml caffeic acid treatment group; 8% butter control group, 8% butter plus 2.5% Echinacea purpura treatment group, 8% butter plus 5% Echinacea purpura treatment group, 8% butter plus 0.075mg/ml caffeic acid treatment group and 8% butter plus 0.15mg/ml caffeic acid treatment group; 13% butter feeding group. Feed Echinacea purpura twice daily and change the caffice acid solution once daily. Another way, we choose the transgenic zebrafish to be animal model, too. Control, 2.5% Echinacea purpura treatment group, 5% Echinacea purpura treatment group, 0.075mg/ml caffeic acid treatment group and 0.15mg/ml caffeic acid treatment group. After feeding butter for one month, we use QPCR to screen the related genes about fatty liver:adiponectin, Apoa1, TNF. The result shows that 8% butter up-regulate of adiponectin, Apoa1 and TNF and it increases fatty liver symptom by cryo-section. So we decide the 8% butter to feed zebrafish for one month and feed Echinacea purpura caffeic acid for 15 days. The result of cryo-section, 0.15mg/ml caffeic acid increase fat in hepatocyte and others are decreased. The QPCR results of gene level about adiponectin, Apoa1, TNFare decreased with all three genes.
中文摘要----------------------------------------------------------------------------5
英文摘要----------------------------------------------------------------------------7
名詞縮寫----------------------------------------------------------------------------7
結果圖--------------------------------------------------------------------------32-51
第一章、 前言---------------------------------------------------------------------10
一、 中草藥-紫錐菊與咖啡酸--------------------------------------------------10
二、 脂肪肝-----------------------------------------------------------------------11
甲、 台灣2008年十大癌症排名
乙、 肝癌的危險因子
丙、人類脂肪肝的形成及機轉
三、 牛油--------------------------------------------------------------------------15
四、 斑馬魚-----------------------------------------------------------------------15
甲、 簡介
乙、 脂質代謝
五、 標的基因--------------------------------------------------------------------21
甲、 adiponetcin
乙、 apoa1
丙、 TNF-
六、 過量Gangkyrin表現於斑馬魚(轉基因魚)及其機轉
第二章、研究目的---------------------------------------------------------------22
第三章、材料與方法------------------------------------------------------------23
一、 斑馬魚-----------------------------------------------------------------------23
二、 飼料製作--------------------------------------------------------------------23
三、 牛油、紫錐菊和咖啡酸的餵食------------------------------------------24
四、 過量Gangkyrin表現於斑馬魚(轉基因魚)
五、 組織染色--------------------------------------------------------------------25
甲、 Hematoxylin and Eosin stain
乙、 Oil Red O stain
六、 即時定量聚合酶連鎖反應-----------------------------------------------26
甲、 Total RNA 抽取
乙、 cDNA 製備
丙、 即時定量聚合酶連鎖反應
第四章、結果---------------------------------------------------------------------29
第五章、討論---------------------------------------------------------------------52
第六章、結論---------------------------------------------------------------------55
參考文獻---------------------------------------------------------------------------56
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