臺灣博碩士論文加值系統

番石榴(Guava, Psidium guajava L.)為熱帶水果的一種，屬於桃金孃科，通常以新鮮的方式食用，也會製成加工食品，例如果汁、糖漿、果醬等。也是台灣重要的經濟作物之一。文獻指出其果實以及葉子具有止瀉，抗菌，抗氧化等功效。本研究以番石榴葉為實驗樣品，探討其不同溶劑之萃取方式對於大鼠正常肝細胞 clone 9 之毒性及其對酒精誘導肝損傷後的保護效果。結果顯示以番石榴萃取物處理 clone 9 細胞24小時，由細胞存活率中發現具有劑量依賴性。根據結果顯示，在細胞存活率試驗上發現水萃取物在400 μg/mL以下之濃度對肝細胞並無毒性以及在60℃熱水萃取物中分別在100 μg/mL以及50 μg/mL對clone 9肝細胞不具有細胞毒性。ALT之活性變化則為丙酮萃取物在200 μg/mL以下而乙醇萃取物在500 μg/mL以下之濃度其ALT在正常值範圍內，在番石榴葉水萃取物以及熱水萃取物組別中對細胞之ALT並無影響。
根據結果顯示應用在clone 9肝細胞之酒精損傷濃度以5％誘導30分鐘做為本實驗之酒精傷肝模式，其中丙酮萃取物以及乙醇萃取物對肝細胞以5％酒精誘導30分鐘後之保護效果在100 μg/mL及50 μg/mL下較為顯著，而水萃取物在200 μg/mL其預防效果較為顯著；熱水萃取物在高濃度下即對細胞有預先保護效果。在ALT（alanine aminotransferase）活性中丙酮萃取物以及乙醇萃取物之預防效果分別在100 μg/mL及50 μg/mL以下之濃度較為顯著。而水萃取物與熱水萃取物分別在50 μg/mL以及200 μg/mL其預防效果較為顯著對於以酒精誘導之肝細胞其保護能力較佳。

Guava (Psidium guajava L.) is a tropical fruit, which belongs to the family Myrtaceae, widely consumed fresh and also processed (beverages, syrup, jams, and ice cream), it is one of the most important economic fruit in Taiwan. Leaves and fruits of guava have been reported to have anti-diarrheal, hypoglycemic, lipid lowering, antibacterial, antioxidant activities. The sample of this study was guava leaf used different solvents extraction, the cytotoxicity of clone 9 liver cells and the hepatoprotective properties of liver injuries induced by alcohol was investigated. When the clone 9 liver cells were treated with guava leaf extracts for 24 hours, the dose-dependent was found. The aqueous extracts was without cytotoxicity in 400 μg/mL, 200 μg/mL, 100 μg/mL and 50 μg/mL, the 60℃ aqueous extracts was without cytotoxicity in 100 μg/mL and 50 μg/mL. According to ALT (alanine aminotransferase) activities, the acetone extracts in 200 μg/mL, ethanol extracts in 500 μg/mL, 400 μg/mL, 200 μg/mL, 100 μg/mL and 50 μg/mL were normal, the aqueous extracts and 60℃ aqueous extracts were without influence.
The clone 9 liver cells cultivated with 5％ alcohol for 30 minutes has been the alcohol injury cell model in this study. According to wst-1 assay and ALT activities, the hepatoprotective properties of acetone extracts, ethanol extracts and aqueous extracts were significant in 100 μg/mL, 50 μg/mL and 50 μg/mL, respectively, 60℃ aqueous extracts had hepatoprotective properties in 200 μg/mL.

目 錄

中文摘要……………………………………………………………….……Ⅰ
英文摘要…………………………………………………………………. Ⅲ
謝誌……………………………………………………………………….. IV
目錄………………………………………………………………………… V
圖表目錄………………………………………………………….………VII
一、前言...........................................................................................................1
二、文獻回顧……………………………………….………………………2
2.1 肝臟簡介……………………………………………………….….3
2.1.1肝臟合成、代謝營養的功能…………………………………………3
2.1.2酒精性肝損傷之作用機制……………………………………….…5
2.2 番石榴之介紹……………………………………………………….13
三、材料與方法…………………………………………………….………18
3.1 細胞來源…………………………………………………….……….18
3.2 實驗材料…………………………………………………….……….18
3.3 實驗藥品…………………………………………………….……….18
3.4儀器設備……………………………………………………………..19
3.5實驗方法……………………………………………………………..20
3.5.1樣品萃取……………………………………………………….20
3.5.2 細胞的培養……………………………………………………...20
3.5.3 細胞株解凍………………………………………………………21
3.5.4細胞冷凍………………………………………………………21
3.5.5 肝細胞形態觀察…………………………………………………21
3.5.6細胞存活率分析-wst-1 assay……………………………………22
3.5.7番石榴萃取物對肝細胞之存活率測定……………………………23
3.5.8酒精對肝細胞之存活率測定………………………………………24
3.5.9番石榴萃取物對酒精傷害肝細胞之預先保護作用………………24
3.5.5肝功能ALT之檢驗………………………………………………24
四、結果與討論……………………………………………………………..26
4.1以酒精誘導clone 9肝細胞損傷試驗……………………………..26
4.2不同溶劑萃取之番石榴葉萃取物對clone 9 之細胞存活率………26
4.3 不同溶劑萃取之番石榴葉萃取物對以酒精誘導損傷之
clone 9肝細胞之細胞存活率..……..……...………….…….………. 26
4.4不同溶劑萃取之番石榴葉萃取物對clone 9 之細胞存活率………38
4.5 不同溶劑萃取之番石榴葉萃取物對以酒精誘導損傷之
clone 9肝細胞之細胞存活率.......…....……….…….……….……….38
5、結論………………………………………………………………………47
6、參考文獻….………………………………………………………………48

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