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研究生:李貞宜
研究生(外文):Chen-Yi Lee
論文名稱:樟芝保肝作用之探討
論文名稱(外文):The Hepatoprotective Effect of Niu-Chang Chih, Antrodia camphorata antihepatoma—in vitro and animal experiments
指導教授:陳健祺林錫璋林錫璋引用關係
指導教授(外文):Jian-Chyi ChenXi-Zhang Lin
學位類別:碩士
校院名稱:南台科技大學
系所名稱:生物科技系
學門:生命科學學門
學類:生物科技學類
論文出版年:2005
畢業學年度:93
語文別:中文
論文頁數:84
中文關鍵詞:樟芝保肝
外文關鍵詞:Antrodia camphorataHepatoprotective
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菇類已經在全世界被廣泛的用來治療疾病有數千年的歷史。其中樟芝為台灣民間之傳統用藥,相信其有某種程度的治療效果,特別是保肝方面。因此本研究藉由MTT assay來評估樟芝萃取物對肝癌細胞(包括:Hep 3B, Hep G2及Alexander cells)生長的抑制效果;另外,以硫代乙醯胺(TAA)誘導大鼠肝纖維化或肝硬化模式作為評估樟芝是否有保肝效果,其功效以Masson trichrome的染色方法評估肝纖維化或肝硬化的程度。此外,也利用體外試驗的方式,以anti-HBsAg assay評估樟芝對anti-HBsAg 的效果。
結果顯示,樟芝水萃物或酒萃物對肝癌細胞Hep 3B, Hep G2及Alexander 細胞增生的抑制率分別為76、77 and 68 %。在動物實驗進行十二週後,TAA成功的誘導極嚴重的肝硬化產生,且在給予樟芝水萃物後能有效的降低肝損傷。此外,在anti-HBsAg assay中,樟芝萃取物也能有效的抑制Alexander cell
所分泌的HBsAg。
此研究結果證明,樟芝水萃物有保護肝臟之能力,此可提供作為在另類醫療中的參考。
Mushrooms have been widely used to prevent and cure diseases for thousand years. The fruit bodies of “niu-chang chih” (Antrodia camphorata) are believed to have certain medical effects in Taiwanese folk medicine, especially for
hepatoprotection.
In this study, the inhibitory effect of Antrodia camphorata on hepatoma cell lines growth (such as Hep 3B, Hep G2 and Alexander cells) was conducted by MTT assay. To evaluate whether there is the hepatoprotective ability of Antrodia camphorata, the rodent model of liver fibrosis, induced by thioacetamide (TAA), has also been set up. The collected livers were stained with Masson trichrome in order to evaluate the degree of liver fibrosis. Furthermore, anti-HBsAg assay was used to investigate the hepatoprotective effect of Antrodia camphorata in reducing the secretion of HBsAg on Alexander cells.
The results demonstrated that there were significantly inhibitory effects of water extract or alcohol extract of mycelium from Antrodia camphorata on Hep 3B, Hep G2 and Alexander cells growth, and the inhibitory rate were 76, 77 and 68% respectively. After twelve week’s TAA-induced, serious liver fibrosis or liver cirrhosis rats have been successfully set up, and the water extract of mycelium from Antrodia camphorata can reduce the liver damage by treated with TAA. Moreover, the secretion of HBsAg on Alexander cells was also significantly inhibited by the water
extract of Antrodia camphorata.
These results suggest that water extract of mycelium from Antrodia camphorata has hepatoprotective ability. The results derived from this study offer some information in alternative medicine.
目 次
中文摘要………………………………………………………………………………. ii
英文摘要…………………………………………………………………………….....iii
目次……………………………………………………………………………………v
表目錄………………………………………………………………………………....vii
圖目錄………………………………………………………………………………...viii
第一章 文獻回顧……………………………………………………………………….1
1-1 肝臟的簡介…………………………………………………………………...1
1-2 肝硬化的簡介………………………………………………………………...2
1-3 硫代乙烯胺(Thioacetamide)的介紹……………………………………..3
1-4 樟芝的介紹…………………………………………………………………...4
1-4-1 分類學………………………………………………………………….....4
1-4-2 外觀與型態特性……………………………………………………….....5
1-4-3 化學成分研究………………………………………………………….....5
1-4-5 安全性評估…………………………………………………………….....6
1-4-6 藥理作用探討………………………………………………………….....7
1-5 水飛薊(Silymarin)的簡介…………………………………………………9
第二章 研究動機與目的……………………………………………………………...11
第三章 材料與研究方法……………………………………………………………..14
3-1 實驗材料…………………………………………………………………….14
3-1-1 樟芝………………………………………………………………………14
3-1-2 水飛薊……………………………………………………………………14
3-1-3 細胞株……………………………………………………………………14
3-1-4 實驗動物…………………………………………………………………14
3-2 實驗藥品………………………………………………………………….....15
3-3實驗套組試劑……………………………………………………………….16
3-4 實驗儀器…………………………………………………………………....16
3-5 實驗方法………………………………………………………………….....17
3-5-1 MTT assay……………………………………………………………......17
3-5-2 Anti-HBsAg assay……………………………………………………......18
3-5-3 Thioacetamide誘導肝硬化model之建立…………………………….20
3-5-4 動物實驗設計…………………………………………………………20
3-5-5 血清測定…………………………………………………………………21
3-5-6 蘇木紫與伊紅(Hematoxylin-Eosin stain)……………………………21
3-5-7 Masson trichrome stain…………………………………………………22
3-5-8 組織病理病變程度的定量……………………………………………23
3-5-9 二維電泳…………………………………………………………………23
3-5-10 統計分析………………………………………………………………..25
第四章 結果…………………………………………………………………………...25
4-1 樟芝水萃物及酒萃物對腫瘤細胞影響…………………………………26
4-2 Silymarin水萃物對腫瘤細胞影響………………………………………28
4-3 樟芝水萃物及酒萃物對HBsAg 的影響……………………………….29
4-4 Thioacetamide誘導肝硬化model之建立………………………………30
4-5 樟芝萃取物對TAA誘導肝損傷之保肝效果…………………………...30
4-6 二維電泳…………………………………………………………………31
第五章 討論…………………………………………………………………………...32
第六章 參考文獻……………………………………………………………………...35
附錄…………………………………………………………………………………….59
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