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研究生:李欣樺
研究生(外文):Hsin-Hua Lee
論文名稱:高氏柴胡抗Diethylnitrosamine誘發大白鼠肝癌之肝臟保護體內活性研究
論文名稱(外文):Preventive Effects of Bupleurum kaoi Against Diethylnitrosamine-induced Hepatocarcinogenesis in Rats
指導教授:顏銘宏顏銘宏引用關係
學位類別:碩士
校院名稱:高雄醫學大學
系所名稱:天然藥物研究所碩士班
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:2006
畢業學年度:94
語文別:中文
論文頁數:90
中文關鍵詞:高氏柴胡Diethylnitrosamine肝癌抗氧化
外文關鍵詞:Bupleurum kaoiDiethylnitrosamineHepatocarcinogenesisAntioxidation
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高氏柴胡為台灣特有種之柴胡,其品質優於現今已知的各種柴胡,目前已知其對大白鼠具有明顯的保肝、抗肝纖維化、抗肝炎的效果。本研究目的為探討台灣產生藥高氏柴胡在化學誘導肝癌動物(大鼠)模式當中的預防作用。在體內試驗中,將Wistar大鼠以腹腔注射投與Diethylnitrosamine (DEN, 200mg/kg)在前兩週給予,接著在第三週~第六週口服給予2-Acetylaminofluorene (2-AAF, 20mg/kg)以誘發肝癌。藥物治療組為高氏柴胡根部的水抽出物(Water extract;BKW)、總多醣體(Polysaccharide-enriched fraction;BKP),以口服給予。實驗結果指出,高氏柴胡於化學誘發肝癌的癌前指標GST-P (Glutathione S-transferase, placental form)、細胞增生指標PCNA (Proliferating cell nuclear antigen)、及進階期的標記TGF-α (Transforming growth factor-alpha)皆有減少的效果,故在免疫組織染色學方面具有化學預防的效果,尤其是BKW。在血清生化值方面,BKW可降低Alkaline phosphatase (AKP)。抗氧化方面,投予BKW、BKP後,顯示兩者在肝均質液內SOD活性中皆會增加,肝臟脂質過氧化產物MDA的量則會降低,而且只有BKW可提升大鼠肝均質液內GSH的量。由上述的結果發現高氏柴胡具有預防由DEN所誘發的化學性肝癌,主要可能是藉由抗氧化作用並清除自由基所造成。
Bupleurum kaoi Liu (Chao et Chuang) is an endemic plant to Taiwan, has been locally used as a hepatoprotective herbs for treating hepatitis, hepatic fibrosis. The preventive effects of water extract (BKW) and polysaccharide-enriched fraction (BKP) from B. kaoi against diethylnitrosamine-induced hepatocarcinogenesis in rats were evaluated in the present study. Male Wistar rats were received 2 doses of diethylnitrosamine (DEN, 200mg/kg, i.p.) 2 weeks apart. One week after the last injection of DEN, the rats were received 20 mg/kg body weight of 2-acetylaminofluorene (2-AAF) by gavage for 3 weeks. Hepatoprotective activity of BKW and BKP were observed in terms of reduced the number of putative preneoplastic altered foci hepatocytes which express with the marker placental form of glutathione-S-transferase (GST-P) and transforming growth factor-alpha (TGF-α). Furthermore, the proliferating cell nuclear antigen (PCNA) was reduced to show the inhibition of foci cell proliferation. In addition, treatment of BKW markedly reduced alkaline phosphatase (AKP) level in rats serum. In view of antioxidation, after treated with BKW and BKP, both of them were showed significantly increased the SOD level and decrease the lipid peroxidation level marker MDA in rats liver homogenates. In addition, only BKW increased GSH level in rats liver homogenates. Our findings suggested that B. kaoi (especially BKW) exhibited preventive activity on the progression phase during DEN-induced hepatocarcinogenesis in rats and the mechanism may due to handle oxidative stress.
中文摘要.................................................5
英文摘要.................................................6

第一章、 序論
第一節 化學誘導肝癌介紹..................................7
第二節 兩階段模式中各階段的標記..........................8
第三節 DEN導致肝臟氧化傷害的機制........................11
第四節 自由基與體內抗氧化機制...........................13
第五節 實驗動物模式設計.................................17
第六節 柴胡(Bupleuri Radix)簡介.........................18
第七節 高氏柴胡(Bupleurum kaoi)介紹.....................19

第二章、材料與方法
第一節 實驗材料的製備...................................21
第二節 實驗藥品與儀器...................................22
第三節 大鼠-體內肝癌誘發之實驗方法......................25
第四節 血清肝功能生化指數的檢測.........................27
第五節 組織病理學觀察...................................28
第六節 肝均質液中抗氧化酵素活性的檢測...................34
第七節 肝均質液中非抗氧化酵素活性的檢測.................38
第八節 肝臟中Malondialdehyde (MDA)含量的測定............40
第九節 統計.............................................43

第三章、結果
第一節 ㄧ般檢測結果.....................................44
第二節 肝功能血清生化值的比較結果.......................45
第三節 組織病理學觀察結果...............................47
第四節 肝均質液中-抗氧化酵素活性比較結果................50
第五節 肝均質液中-非抗氧化酵素活性比較結果..............51
第六節 肝臟中Malondialdehyde (MDA)含量的測定結果........52

第四章、討論............................................53

第五章、結論............................................60

附錄(表)................................................61

附錄(圖)................................................68

參考文獻................................................83

學會發表論文............................................90
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