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研究生:張依如
研究生(外文):I-JU, Chang
論文名稱:鞣花酸對乙醯胺酚誘發大鼠肝損傷之抗氧化及護肝作用
指導教授:盧義發盧義發引用關係
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
論文種類:學術論文
論文出版年:2010
畢業學年度:98
語文別:中文
中文關鍵詞:鞣花酸乙醯胺酚抗氧化肝損傷
外文關鍵詞:ellagic acidacetaminophenantioxidanthepatotoxicity
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許多研究指出過量服用乙醯胺酚 (acetaminophen, N-acetyl-p-aminophenol, APAP) 的鎮熱、止痛藥物會造成肝臟損傷。鞣花酸(ellagic acid, EA)為酚類化合物,具有抗致突變、抗腫瘤及抗氧化活性。本研究欲探討鞣花酸是否能改善APAP 所誘發之肝損傷及對體內抗氧化與脂質代謝之影響。將 32 隻 Sprague-Dawley 大鼠分為四組:分別為控制組 (腹腔注射 50% 丙二醇/生理食鹽水)、 APAP組 ( 500 mg /kg BW),實驗組 (注射 APAP 0.5小時後,給予EA 50 mg /kg BW) 及正控制組 (給予 N-acetylcysteine 1g /kg BW),24小時後犧牲,取血液及肝臟進行分析。結果發現,EA 可降低肝損傷大鼠血清中天門冬胺酸轉胺酶 (aspartate aminotransferase)、丙胺酸轉胺酶 (alanine aminotransferase)、鹼性磷酸酶 (alkaline phosphatase) 及伽瑪麩胺醯轉移酶 (gamma-glutamyl transpeptidase) 活性;抗氧化酵素活性方面,EA 可增加過氧化氫酶活性,降低脂質過氧化物生成,並能增加肝臟中總抗氧化能力及麩胱甘肽 (glutathione, GSH) 含量,提高 GSH/GSSG 的比值,對於超氧歧化酶、麩胱甘肽過氧化酶及麩胱甘肽硫轉移酶等酵素活性則無顯著影響。另外,組織切片結果顯示 EA 可減輕 APAP 所引起的病理組織改變。此外,利用高效液相層析儀分析國內桃園一號及三號草莓品種中鞣花酸含量,每 100 g 草苺鮮重約含 20 mg 鞣花酸。以上結果,推測 EA 可緩解APAP 所造成大鼠的肝損傷,並提升其體內抗氧化能力。
目錄 頁次

中文摘要…………………………………………………………………………… I
英文摘要…………………………………………………………………………… II
致謝…………………………………………………………………………………III
目錄………………………………………………………………………………IV
表目錄……………………………………………………………………………VII
圖目錄……………………………………………………………………………VIII
附圖目錄………………………………………………………………………VIII
壹、 緒言……………………………………………………………………………1
貳、 文獻回顧………………………………………………………………………2
一、 自由基與氧化壓力………………………………………………………… 2
(一) 自由基與活性氧………………………………………………………… 2
(二) 自由基與活性氧來源…………………………………………………… 2
(三) 氧化壓力與氧化傷害…………………………………………………… 3
二、 體內抗氧化防禦系統………………………………………………………4
(一) 酵素性抗氧化防禦系統………………………………………………… 4
(二) 非酵素性抗氧化防禦系統……………………………………………… 6
三、 體內解毒系統……………………………………………………………… 9
(一) 細胞色素 P450 系統.................................... 9
(二) 麩胱甘肽硫轉移酶...................................... 10
四、 肝臟………………………………………………………………...... 10
(一) 肝炎……………………………………………………………………... 11
(二) 肝發炎指數……………………………………………………………... 11
五、 乙醯胺酚…………………………………………………………………… 12
(一) 乙醯胺酚之歷史背景與臨床應用……………………………………… 12
(二) 乙醯胺酚之肝毒性與代謝途徑………………………………………… 13
(三) 乙醯胺酚之中毒劑量................................... 14
(四) N-乙醯半胱胺酸...................................... 15
六、 鞣花酸……………………………………………………………………… 15
(一) 鞣花酸的基本性質……………………………………………………… 15
(二) 鞣花酸之生理活性……………………………………………………… 17
(三) 鞣花酸之安全性………………………………………………………… 18
七、 台灣草莓品種的變遷……………………………………………………… 19
八、 台灣草莓成分分析………………………………………………………….19
參、 材料與方法…………………………………………………………………… 20
一、 藥品與器材………………………………………………………………… 20
(一) 分析用試藥……………………………………………………………… 20
(二) 儀器設備………………………………………………………………… 21
二、 實驗設計與架構……………………………………….…………………… 22
實驗I:分析台灣主要草莓品種中鞣花酸含量………….…………………..… 22
(一) 實驗目的……………………………………….………………...……… 23
(二) 實驗材料……………………………………….………………...……… 23
(三) 實驗方法……………………………………….………………...……… 23
1. 萃取條件……………………..…………….………………...……… 23
2. 分析條件……………………..…………….………………...……… 23
3. 統計分析……………………..…………….………………...……… 23
實驗 Ⅱ:鞣花酸與N-acetylcysteine對 APAP 誘發肝毒性之緩解效果........ 24
(一) 實驗目的……………………………………….………………...……… 25
(二) 實驗材料……………………………………….………………...……… 25
(三) 實驗方法……………………………………….………………...……… 25
1. 動物飼養條件與分組…………………….….……………...……… 25
2. 實驗時間點設計………………………….….……………...……… 25
3. 樣本收集與處理…………………….….……………...........……… 26
4. 樣本分析…………………….….…………………………...……… 26
5. 統計分析………………………………….….……………...……… 31
肆、 結果
一、 草莓中鞣花酸含量分析………………….……………………………….. 32
二、 對體重與肝臟重量之影響……………….……………………………….. 32
三、 對血清肝功能指標之影響……………….……………………………….. 32
四、 對血清及肝臟中抗氧化物質之影響…….……………………………….. 32
五、 對血清及肝臟中總抗氧化力與脂質過氧化之影響……………………... 33
六、 肝臟組織病理變化之影響………………………………….…………….. 34
伍、 討論………………………………………………………………………...… 35
陸、 結論………………………………………………………………………...… 42
柒、 參考文獻………….………………………………………………………….. 57

表目錄 頁次

表一、各植物中鞣花酸的含量.................................... 17
表二、台灣草莓中鞣花酸含量及含水率............................. 43
表三、不同抗氧化劑對大鼠體重及肝重之影響........................ 44
表四、不同抗氧化劑對乙醯胺酚誘發肝損傷的大鼠肝功能指標之影響...... 45
表五、不同抗氧化劑對乙醯胺酚誘發肝損傷的大鼠血清 catalase 及 SOD
之影響................................................ 46
表六、不同抗氧化劑對乙醯胺酚誘發肝損傷的大鼠肝臟 catalase 及 SOD
之影響................................................ 47
表七、不同抗氧化劑對乙醯胺酚誘發肝損傷的大鼠肝臟 GPx, GSH, GSSG 及
GSH/GSSG 比值之影響.................................... 48
表八、不同抗氧化劑對乙醯胺酚誘發肝損傷的大鼠血清總抗氧化力
(TEAC) 及脂質過氧化物 (TBARS) 之影響.................... 49
表九、不同抗氧化劑對乙醯胺酚誘發肝損傷的大鼠肝臟總抗氧化力
(TEAC) 及脂質過氧化物 (TBARS) 之影響.................... 50

圖目錄 頁次

圖一、乙醯胺酚代謝機制...................................... 14
圖二、鞣花酸結構式......................................... 16
圖三、鞣花酸標準品之 HPLC 分析圖譜...........................51
圖四、桃園一號草莓樣品之 HPLC 分析圖譜........................52
圖五、桃園三號草莓樣品之 HPLC 分析圖譜........................53
圖六、草莓樣品及樣品添加鞣花酸標準品後之 HPLC 分析圖譜…………….54
圖七、不同抗氧化劑對乙醯胺酚誘發肝損傷的大鼠肝臟 GST 之影響.... 55
圖八、大鼠肝臟組織切片圖................................... 56

附圖目錄 頁次

附圖一、不同抗氧化劑對乙醯胺酚誘發肝損傷的大鼠肝功能指標之影響.... 74
附圖二、不同抗氧化劑對乙醯胺酚誘發肝損傷大鼠血清及肝臟 SOD 與
catalase 之影響..................................... 75
附圖三、不同抗氧化劑對乙醯胺酚誘發肝損傷的大鼠肝臟 GPx, GSH 與GSSG
之影響............................................. 76
附圖四、不同抗氧化劑對乙醯胺酚誘發肝損傷大鼠血清及肝臟總抗氧化力
與脂質過氧化之影響................................... 77
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