臺灣博碩士論文加值系統

本研究旨探討貯存溫度與時間對石蓴酒組成份及抗氧化活性之影響，以及餵食石蓴酒對大鼠脂質代謝及抗氧化能力之影響。石蓴酒分別放置於4℃、10℃及15℃貯存7個月後，隨貯酒時間的增加，石蓴酒總酚類化合物含量、抑制血紅素誘導亞麻油酸過氧化能力、還原力及總抗氧化能力均隨之增加。剛發酵完之石蓴酒揮發性成份約十一種，貯存七個月後，貯存於4℃之石蓴酒揮發性成份增至十二種，但於10℃及15℃貯存之石蓴酒揮發性成份減至十種。所有酒液中以ethyl octanoate 含量最高。SD大鼠餵食食用酒精 (2 g酒精/公斤體重) 12週，血漿中三酸甘油酯 (TG) 和總膽固醇 (TC) 含量明顯增加，分別為控制組之123.6% 與110.8%。血漿中高密度脂蛋白膽固醇(HDL-C) 含量則下降13%。另外，血漿和肝臟丙二醛 (MDA) 含量明顯高於控制組，分別為控制組之151.7% 和125.8%。餵食同等酒精劑量石蓴酒之SD大鼠其血漿TG、TC、HDL-C和MDA含量與控制組均無顯著差異。餵食酒精之SD大鼠肝臟超氧岐化酶 (SOD)、觸酶 (catalase)、肝臟麩胱甘肽過氧化酶 (GPx) 和麩胱甘肽還原酶活性也顯著低於控制組，餵食石蓴酒之SD大鼠與控制組則無顯著差異。餵食石蓴酒之SD大鼠肝臟麩胱甘肽 (GSH) 與氧化態麩胱甘肽 (GSSG) 比值明顯低於控制組，但酒精組之SD大鼠GSH/GSSG比值均顯著低於石蓴酒組。大鼠肝臟組織切片觀察發現，餵食食用酒精之SD大鼠肝臟以Oil Red O和Hematoxylin染劑染色後，在肝細胞周圍有明顯的紅色脂肪滴，石蓴酒組之SD大鼠則較輕微。另以Hematoxylin和Eosin染劑染色觀察大鼠肝臟細胞形態之變化，發現
酒精組大鼠受損較石蓴酒組及控制組明顯。

　　The aims of this research are to investigate the changes in flavor and antioxidant activity of Ulva wine during storage at various temperatures, and to evaluate its antioxidant activity in rats. The total phenolic content and antioxidative activity including reducing activity, inhibition activity for hemoglobin-induced oxidation of linoleic acid, and total antioxidant status were increased during Ulva wine aging at 4℃, 10℃ or 15℃. Eleven volatile compounds were detected by GC-MS for fresh fermented Ulva wine. One more volatile compound was detected after storage at 4℃ for 7 months; while at the same time 1 volatile compounds were vanished for Ulva wine stored at 10℃ or 15℃. Ethyl octanoate was the dominant volatile compound for all Ulva wine aged at 4℃, 10℃ or 15℃. After being fed with ethanol at dose of 2 g ethanol/kg B.W. for 12 weeks, the plasma triacylglycerol (TG) and total cholesterol (TC) of SD rats were increased by 123.6% and 110.8%, respectively, compared to control group. Their malondialdehyde (MDA) contents in plasma and liver were icreased by 151.7% and 125.8%, respectively; while high density lipoprotein cholesterol (HDL-C) in plasma was significantly decreased. However, there were no significantly different for TG、TC、HDL-C and MDA between the control group rats and rats fed with Ulva wine for 12 weeks. The activities of superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx) and glutathione reductae were significantly decreased, compared to those of control. While there were no significantly differerent in those 4 antioxidative enzymes between rats fed with Ulva wine and control. By counterstaining with Oil Red O and Hematoxyline more lipid in liver section was observed for rats fed with ethanol. Liver lesion was also observed for rats exposed to ethanol.

目錄
中文摘要------------------------------------------------------------------------------------------I
英文摘要----------------------------------------------------------------------------------------III
壹、 前言------------------------------------------------------------------------------------------1
貳、 文獻整理------------------------------------------------------------------------------------2
一、 石蓴------------------------------------------------------------------------------------------2
1. 成份-------------------------------------------------------------------------------------2
2. 抗發炎----------------------------------------------------------------------------------3
3. 抗微生物活性-------------------------------------------------------------------------4
4. 降血脂----------------------------------------------------------------------------------4
5. 其它功用-------------------------------------------------------------------------------5
二、 酒之風味成份-----------------------------------------------------------------------------6
1. 醇類-------------------------------------------------------------------------------------6
2. 酯類-------------------------------------------------------------------------------------7
3. 含硫化合物----------------------------------------------------------------------------7
4. 其它成份-------------------------------------------------------------------------------8
三、 熟成作用------------------------------------------------------------------------------------8
1. 溫度-------------------------------------------------------------------------------------9
2. 時間-------------------------------------------------------------------------------------9
3. 容器------------------------------------------------------------------------------------10
四、 酒精對生物體之影響-------------------------------------------------------------------10
1. 酒精代謝------------------------------------------------------------------------------11
2. 氧化壓力之產生---------------------------------------------------------------------11
3. 酒精性脂肪肝之形成---------------------------------------------------------------12
4. 適量飲酒之好處---------------------------------------------------------------------14
5. 酒精與抗氧化------------------------------------------------------------------------14
參、 實驗設計----------------------------------------------------------------------------------16
肆、 材料與方法------------------------------------------------------------------------------17
一、材料---------------------------------------------------------------------------------------17
1. 原料-----------------------------------------------------------------------------------17
2. 培養基---------------------------------------------------------------------------------17
3. 藥品------------------------------------------------------------------------------------17
4. 動物------------------------------------------------------------------------------------18
二、儀器設備-----------------------------------------------------------------------------------19
三、實驗方法-----------------------------------------------------------------------------------20
1. 石蓴酒之釀製------------------------------------------------------------------------20
1.1 石蓴前處理----------------------------------------------------------------------20
1.2 菌株保存-------------------------------------------------------------------------20
1.3 菌株活化-------------------------------------------------------------------------20
1.4 發酵酒液-------------------------------------------------------------------------21
2. 貯酒試驗------------------------------------------------------------------------------21
2.1 貯酒過程之成份分析----------------------------------------------------------21
2.1.1 酵母菌菌數分析-------------------------------------------------------21
2.1.2 酒精度分析-------------------------------------------------------------22
2.1.3 總酸分析----------------------------------------------------------------22
2.1.4 殘糖測定----------------------------------------------------------------22
2.2 抗氧化活性測定---------------------------------------------------------------23
2.2.1 總酚含量分析----------------------------------------------------------23
2.2.2 抑制血紅素催化亞麻油酸過氧化之能力-------------------------23
2.2.3 還原力測定-------------------------------------------------------------24
2.2.4 Trolox 當量抗氧化能力----------------------------------------------24
2.3 揮發性成份測定----------------------------------------------------------------25
2.3.1 固相微量萃取器前處理----------------------------------------------25
2.3.2 石蓴酒揮發性成份分析----------------------------------------------25
2.3.3 氣相層析儀之條件----------------------------------------------------26
2.4 品評試驗------------------------------------------------------------------------27
3. 動物試驗------------------------------------------------------------------------------27
3.1 血漿分析------------------------------------------------------------------------28
3.1.1 總膽固醇之測定-------------------------------------------------------28
3.1.2 三酸甘油酯之測定----------------------------------------------------28
3.1.3 脂蛋白-膽固醇之測定------------------------------------------------28
3.1.4 血中尿素氮之測定----------------------------------------------------29
3.1.5 肌酸酐之測定----------------------------------------------------------29
3.1.6 血漿麩酸草酸轉胺酶-------------------------------------------------29
3.1.7 血漿麩酸丙酮酸轉胺酶----------------------------------------------29
3.1.8 TBARS濃度測定------------------------------------------------------30
3.1.9 血漿總抗氧化狀態----------------------------------------------------30
3.1.10 血漿一氧化氮含量----------------------------------------------------30
3.2 肝臟分析-------------------------------------------------------------------------31
3.2.1 肝臟脂質測定----------------------------------------------------------31
3.2.2 TBARS 濃度測定-----------------------------------------------------31
3.2.3 觸酶活性測定----------------------------------------------------------31
3.2.4 超氧岐化酶活性測定-------------------------------------------------32
3.2.5 麩胱甘肽與氧化態麩胱甘肽測定----------------------------------32
3.2.6 麩胱甘肽過氧化酶活性測定----------------------------------------33
3.2.7 麩胱甘肽還原酶活性測定-------------------------------------------34
3.2.8 蛋白質含量測定------------------------------------------------------34
3.3 病理組織切片-----------------------------------------------------------------34
3.3.1 石臘組織切片---------------------------------------------------------34
3.3.2 組織切片之固定------------------------------------------------------35
3.3.3 石臘包埋---------------------------------------------------------------35
3.3.4 石臘切片---------------------------------------------------------------35
3.3.5 切片染色---------------------------------------------------------------35
3.3.6 冷凍切片---------------------------------------------------------------36
3.3.6.1組織切片---------------------------------------------------------------36
3.3.6.2 脂肪組織之染色------------------------------------------------------36
4. 統計分析-----------------------------------------------------------------------------36
伍、 結果與討論-------------------------------------------------------------------------------37
一、 石蓴酒貯酒過程成份之變化----------------------------------------------------37
二、 石蓴酒貯酒過程對抗氧化活性之影響----------------------------------------37
三、 石蓴酒貯酒過程揮發性成份之變化-------------------------------------------38
四、 石蓴酒對大鼠體重、肝臟及腎臟比例之變化--------------------------------39
五、 石蓴酒對大鼠血漿生化值之影響----------------------------------------------40
六、 石蓴酒對大鼠血漿一氧化氮含量及肝臟脂質之影響----------------------41
七、 石蓴酒對大鼠脂質過氧化之影響----------------------------------------------43
八、 石蓴酒對大鼠體內抗氧化活性之影響----------------------------------------44
九、 石蓴酒對大鼠肝臟組織病理之影響-------------------------------------------48
陸、 結論----------------------------------------------------------------------------------------50
柒、 參考文獻----------------------------------------------------------------------------------51














圖表目錄
圖一、不同溫度貯存過程石蓴酒中總酚含量之變化--------------------62
圖二、不同溫度貯存過程石蓴酒中抑制血紅素誘導亞麻油酸過氧化之變化------------------------------------------------------------------------62
圖三、不同溫度貯存過程石蓴酒還原力之變化-------------------------63
圖四、不同溫度貯存過程石蓴酒中總抗氧化能力之變化-------------63
圖五、SD大鼠分別餵食蒸餾水(控制組)、石蓴酒及酒精12週腹腔解剖觀察腹腔脂肪變化--------------------------------------------------64
圖六、SD大鼠分別餵食蒸餾水(控制組)、石蓴酒及酒精12週後血漿中一氧化氮濃度---------------------------------------------------------65
圖七、SD大鼠分別餵食蒸餾水(控制組)、石蓴酒及酒精12週後肝臟總膽固醇和三酸甘油酯之含量--------------------------------------66
圖八、SD大鼠分別餵食蒸餾水(控制組)、石蓴酒及酒精12週後血漿總抗氧化狀態------------------------------------------------------------67
圖九、SD大鼠分別餵食蒸餾水(控制組)、石蓴酒及酒精12週後以病理組織切片觀察肝臟脂肪滴之變化 (Oil Red O stain) ----------------68
圖十、SD大鼠分別餵食蒸餾水(控制組)、石蓴酒及酒精12週後以病理組織切片觀察肝臟脂肪滴之變化 (Oil Red O & hematoxylin stain) -----------------------------------------------------------------------69

圖十一、SD大鼠分別餵食蒸餾水(控制組)、石蓴酒及酒精12週後以病理組織切片觀察肝組織之變化 (H&E stain) ------------------------70
表一、不同溫度貯存過程石蓴酒酵母菌數、酒精度、總酸、pH和殘糖之變化------------------------------------------------------------------71
表二、不同溫度貯存過程石蓴酒揮發性成份之變化--------------------72
表三、不同溫度貯存石蓴酒之品評分析-----------------------------------73
表四、SD大鼠分別餵食蒸餾水(控制組)、石蓴酒及酒精12週後體重、腎臟和肝臟重量之變化------------------------------------------------74
表五、SD大鼠分別餵食蒸餾水(控制組)、石蓴酒及酒精12週後血漿中三酸甘油酯、總膽固醇、高密度脂蛋白-膽固醇、低密度脂蛋白-膽固醇、血中尿素氮、肌酸、血清麩酸草酸轉胺酶和血清麩酸丙酮酸轉胺酶之含量 ----------------------------------------------75
表六、SD大鼠分別餵食蒸餾水(控制組)、石蓴酒及酒精12週後血漿和肝臟脂質過氧化值(以丙二醛量表示) --------------------------75
表七、SD大鼠分別餵食蒸餾水(控制組)、石蓴酒及酒精12週後肝臟抗氧化酵素超氧岐化酶和觸酶之活性-----------------------------76
表八、SD大鼠分別餵食蒸餾水(控制組)、石蓴酒及酒精12週後肝臟麩胱甘肽過氧化酶、麩胱甘肽還原酶之活性，及還原態麩胱甘肽、氧化態麩胱甘肽之含量------------------------------------------76

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