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研究生:劉家瑜
研究生(外文):Liu, Jiayu
論文名稱:人工段木培養之牛樟芝子實體萃取物對半乳糖胺誘發大鼠 急性肝損傷之保護作用
論文名稱(外文):Hepatoprotective effects of Taiwanofungus camphorates fruiting bodies extract from timber cultivated by D-galactosamine-induced liver damage in rats
指導教授:盧義發盧義發引用關係
指導教授(外文):Lu, Yi-Fa
口試委員:蔡震壽黃惠宇林万登劉沁瑜
口試日期:2012-07-06
學位類別:碩士
校院名稱:輔仁大學
系所名稱:營養科學系
學門:醫藥衛生學門
學類:營養學類
論文出版年:2012
畢業學年度:100
語文別:中文
論文頁數:92
中文關鍵詞:牛樟芝子實體半乳糖胺護肝作用抗發炎抗氧化三萜類化合物
外文關鍵詞:Taiwanofungus camphorates fruiting bodiesD-galactosaminehepatoprotective effectsanti-inflammatorytriterpenes
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肝臟為人體主要解毒、代謝器官,肝臟受到急性病毒感染、藥物毒性或其他原因,皆可能造成急性肝衰竭,其死亡率高達八成。野生牛樟芝為台灣特有蕈菇類,富含三萜類、多醣體和多酚類等。本研究以人工段木培養牛樟芝子實體為材料,探討給予大鼠不同濃度之酒精萃取物,是否對於半乳糖胺 (D-GalN) 所誘發之急性肝損傷具有預防與保護作用。將八週齡之Wistar雄性大鼠分為6組,連續七天經口管餵給予不同劑量牛樟芝萃取物 (0.25、0.75 mg/day) 及常用保肝劑Silymarin,於第7天最後一次管餵4小時後,以腹腔注射給予D-GalN誘導急性肝損傷,24小時後犧牲。結果顯示,牛樟芝高劑量組可降低肝損傷大鼠血清中天門冬胺酸轉胺酶 (AST) 及丙胺酸轉胺酶 (ALT),並且降低血清中腫瘤壞死因子-α (TNF-α),提升肝臟超氧歧化酶 (SOD) 活性,降低血清脂質過氧化程度 (TBARS) ,達到減緩發炎、提升抗氧化能力及預防肝臟嚴重損傷。此外,在體外抗氧化試驗發現以熱水萃取物具有最佳的體外抗氧化能力,但產率與總多酚含量最低,推測人工段木培養牛樟芝子實體強抗氧化活性來自於水溶性之粗多醣成份。利用高效液相層析儀分析人工段木牛樟芝子實體與野生牛樟芝子實體具有相似成分,亦為三萜類的良好來源。
Liver is the major detoxification and metabolic organ in human body. The acute liver failure (Fulminant hepatic failure, FHF) were caused by viral infection, drug toxicity or other reasons. Recently, FHF carried a high mortality rate of 80%. Taiwanofungus camphorates (TC), riched in physiologically active substances, such as triterpenes, polysaccharides, polyphenols and others is a fungus that only grows in Taiwan. The aim of this study is to investigate the hepatoprotective effects of TC fruiting bodies extract from timber cultivated by D-galactosamine-induced liver damage in rat, and explore the potential anti-inflammatory and anti-oxidative properties. The rats were randomly divided into six groups (n = 6 each). In experimental group, TC extract (0.25, 0.75 mg/day) or hepatoprotective agent commonly used Silymarin (25 mg / kg BW) was administered orally once a day for 1 week, and D-GalN injected 4 hr after last gavage. The rats were sacrificed after 24 hr of D-GalN treatment. The results showed that the high dose of TC extracts had significantly decreased levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), tumor necrosis factor-alpha (TNF-alpha) and thiobarbituric acid reactive substances (TBARS) in serum, and higher levels of superoxide dismutase (SOD) activity in liver compared with the negative control group. In addition, hot water extracts from timber cultivated TC fruiting bodies were better anti-oxidant capacity test in vitro than ethanol extracts , but the yield and total phenolic content of hot water extracts were the lowest one. So we speculated that the antioxidant activity of TC hot water extracts from the water-soluble crude polysaccharide composition. The high performance liquid chromatogm of TC fruiting bodies extracts from timber cultivated were similar to wild TC fruiting bodies extracts, and showed a good source of TC triterpenes.
中文摘要………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………I
英文摘要………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………II
目錄………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………III

壹、緒言………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………1
貳、文獻回顧……………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………3
一、牛樟芝介紹…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………3
(一)來源………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………3
(二)牛樟芝成分探討…………………………………………………………………………………………………………………………………………………………………………………………………………………………………5
(三)牛樟芝的生理活性………………………………………………………………………………………………………………………………………………………………………………………………………………………………7
二、肝臟生理機能與損傷………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………10
(一)肝臟的功能…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………11
(二)肝臟抗氧化酵素系統………………………………………………………………………………………………………………………………………………………………………………………………………………………11
(三)發炎反應………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………13
(四)肝發炎指數…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………14
(五)肝臟損傷模式………………………………………………………………………………………………………………………………………………………………………………………………………………………………………15
(六)常用保肝劑-水飛薊素………………………………………………………………………………………………………………………………………………………………………………………………………………………19
參、材料與方法…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………20
一、實驗目的與設計架構………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………20
二、試驗材料……………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………21
(一)牛樟芝子實體………………………………………………………………………………………………………………………………………………………………………………………………………………………………………22
(二)分析試藥………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………22
三、儀器設備……………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………23
四、實驗方法………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………24
(一)實驗I ……………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………24
(二)實驗II……………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………29
五、統計分析………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………33
肆、結果 ……………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………34
一、實驗I 34
(一)大鼠體重變化及肝臟重量…………………………………………………………………………………………………………………………………………………………………………………………………………………34
(二)大鼠血清肝功能指標…………………………………………………………………………………………………………………………………………………………………………………………………………………………34
(三)大鼠血清中腫瘤壞死因子…………………………………………………………………………………………………………………………………………………………………………………………………………………35
(四)大鼠血清中介白素-10………………………………………………………………………………………………………………………………………………………………………………………………………………………35
(五)大鼠肝臟中抗氧化酵素之活性…………………………………………………………………………………………………………………………………………………………………………………………………………35
(六)大鼠血清及肝臟脂質過氧化程度……………………………………………………………………………………………………………………………………………………………………………………………………36
(七)大鼠血清及肝臟總抗氧化能力…………………………………………………………………………………………………………………………………………………………………………………………………………36
二、實驗II ………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………37
(一)人工段木培養牛樟芝子實體在不同溶劑之萃取率………………………………………………………………………………………………………………………………………………………………………37
(二)人工段木培養牛樟芝子實體之粗三萜類與總多醣含量………………………………………………………………………………………………………………………………………………………………37
(三)人工段木培養牛樟芝子實體指標化合物比對………………………………………………………………………………………………………………………………………………………………………………37
(四)人工段木培養牛樟芝子實體萃取物體外抗氧化能力比較…………………………………………………………………………………………………………………………………………………………38
伍、討論 ……………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………40
陸、結論 ……………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………50
柒、參考文獻 ……………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………74



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