臺灣博碩士論文加值系統

本研究採用食品工業發展研究所菌種中心所生產之牛樟芝菌絲體發酵過濾液(filtrate of fermented mycelia of Antrodia camphorata, FFMA)為實驗樣品，探討其對四氯化碳誘導大白鼠慢性肝損傷之影響。本實驗共分為六組，為正常對照組(未經四氯化碳處理)，其餘皆經四氯化碳處理組，分別為負對照組(僅投予四氯化碳之處理組)、正對照組(Silymarin之處理組)及三組實驗組。三組實驗組分別為低、中、高劑量，3、12.4及24.8 ml/kg BW/day FFMA處理組。大白鼠於四氯化碳投予前一週每日一次經口投予FFMA，直到實驗終了。實驗結果顯示，在第八週的血清生化值glutamic oxaloacetic transaminase (GOT)、glutamic pyruvic transaminase (GPT)方面，三組FFMA實驗組及正對照組的GOT、GPT值皆較負對照組低且有顯著性差異(P<0.05)。在第八週血清中triglyceride (TG)、cholesterol (Chol.)值方面，中、高劑量FFMA實驗組與正對照組之TG、Chol.皆較負對照組為高且有顯著性差異(P<0.05)。在肝臟脂質過氧化方面，三組FFMA實驗組與正對照組與負對照組比較皆有顯著性下降的現象(P<0.05)；血漿脂質過氧化方面，高劑量FFMA實驗組之TBARS值較負對照組為低且有顯著性差異(P<0.05)。在肝臟抗氧化及解毒代謝酵素活性方面，經中、高劑量FFMA處理之實驗組之glutathione peroxidase (GPx)、glutathione reductase (GRd)及glutathione S-transferase (GST)活性均顯著較負對照組高(P<0.05)；紅血球中抗氧化與解毒代謝酵素GPx、GRd及GST活性方面，高劑量FFMA實驗組皆較負對照組有顯著性上升的現象(P<0.05)。在肝臟GSH影響方面，中、高劑量FFMA實驗組GSH含量與負對照組比較有顯著性的上升的情形(P<0.05)；紅血球中GSH含量方面，FFMA實驗組與負對照組比較有顯著性的上升的情形(P<0.05)。而中、高劑量FFMA實驗組之肝臟superoxide dimutase (SOD)活性與負對照組比較亦有顯著性上升的情形(P<0.05)，三組FFMA實驗組之肝臟Catalase (CAT)活性與負對照組比較則有上升的情形，但未達顯著性差異。另外，而高劑量FFMA實驗組之紅血球SOD及Catalase活性與負對照組比較，皆有顯著性的上升的情形(P<0.05)。在肝臟TG及Chol.值方面，FFMA實驗組之TG值均較負對照組為低且有顯著性差異(P<0.05)；正對照組及FFMA實驗組之Chol.值與負對照組比較雖有下降情形，但無顯著性差異。在第八週經四氯化碳誘發慢性肝損傷之大白鼠hydroxyproline (HYP)值皆顯著性高於正常對照組(P<0.05);三組不同劑量FFMA實驗組之HYP值皆較負對照組為低且有顯著性差異(P<0.05)。肝臟組織病理學觀察方面，三組FFMA實驗組之肝變性程度均較負對照組顯著性降低 (P<0.05)，又以中、高劑量效果最明顯(P<0.01)。在肝纖維化方面，三組FFMA實驗組較負對照組皆有顯著性降低纖維化程度。因此適量的牛樟芝菌絲體發酵過濾對肝臟生理機能性具有正面的影響，可降低肝纖維化及變性程度，增加肝臟與紅血球中GSH含量及增加抗氧化酵素活性，降低肝臟脂質過氧化程度，而有利於體內氧化壓力的下降。

中文摘要……………………………… ………………………Ⅳ
前言 …..…………………………………………………1
文獻回顧………………………………………………………3
一、牛樟芝…………………………………………………… 3
(一) 牛樟芝簡介……………………………………………3
(二)成份分析……………………………………………………5
(三)生理機能研究………………………………………………7
(四)安全毒性研究………………………………………………8
二、四氯化碳誘導慢性肝損傷 ……………………….................11
三、自由基與抗氧化……………………………………………….14
材料與方法…………………………………… ………………………19
一、實驗材料……………………………………………………….20
二、實驗動物……………………………….……………………....20
三、實驗分組……………………………………………………….20
四、四氯化碳誘導慢性肝損傷…………………………………….21
(一)投予方法…………………………………………………...21
(二)實驗設計…………………………………………………...21
(三)犧牲當天實驗流程……………………………………… .22
五、樣品製備……………………………………………………….22
(一)肝臟………………………………………………………..22
(二)血液………………………………………………………..22
六、實驗樣品分析………………………………………………….23
(一)血液生化值檢測……………………………………………23
(二)肝臟麩胱甘（glutathione, GSH）含量之分析………….23
(三)紅血球中麩胱甘（glutathione, GSH）含量之分析…….25
(四)肝臟脂質過氧化之分析…………………………………….26
(五)肝臟與紅血球內麩胱甘過氧化（glutathione peroxidase,
GPx）活性之分析…………………………………………….27
(六)肝臟與紅血球內麩胱甘還原（glutathione reductase, GRd）活性之分析…………………………………………….28
(七)肝臟與紅血球內麩胱甘硫轉移（glutathione S-transferase, GST）活性之分析……………………………..29
(八)肝臟與紅血球中超氧化物岐化（superoxide dismutase, SOD）活性之分析…………………………………………….30
(九)肝臟與紅血球中過氧化氫（catalase, CAT）活性之分析………………………………………………………………31
(十)蛋白質定量分析………..........................32
(十一)組織病理學觀察…. …………………………………32
(十二)肝臟膠原蛋白含量之定量分析……… ………………33
七、統計分析…………………………………………………34.
結 果………………………………………….. …………37
一、肝臟重/體重比....................................................37
二、平均體重變化………………………………………………….37
三、在血清生化值………………………………………………….38
四、肝臟脂質過氧化……………………………………………….42
五、肝臟之抗氧化與解毒代謝酵素活性………………………….42
六、肝臟之麩胱甘（GSH/GSSG）活性………………………….43
七、肝臟其他抗氧化酵素活性…………………………………….44
八、肝臟TG及Chol.值……………………………………………45
九、肝臟Hydroxyproline（HYP）含量…………………………..45
十、肝臟組織病理學觀察………………………………………….46
十一、血漿脂質過氧化…………………………………………….46
十二、紅血球抗氧化與解毒代謝酵素活性……………………….47
十三、紅血球麩胱甘（GSH/GSSG）…………………………….48
十四、紅血球其他抗氧化酵素活性……………………………….49
討 論………………………………………………………………50
結 論…………………………………………………… .… …58
表………………………………………………………………………59
圖………………………………………………………… … ………35
參考文獻………………………………………………………………75.

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