- 學位論文
1. 在四氯化碳誘導大鼠肝損傷模式下中草藥複方對於護肝的效果之探討 2. 探討巴西洋菇發酵產物之乙醇萃取物對接種肝癌細胞株Hep3B之裸鼠的影響
1. Hepatoprotective Effect of Herbal Complex on Liver Damage Induced by Carbon Tetrachloride in Rats 2. Effects of the Ethanolic Extract of the Fermentation Product of Agaricus blazei on Hepatocellular Carcinoma (Hep 3B Cells)-implanted in Nude Mice
摘要
1. 慢性疾病自2005年後已逐漸成為世界人口死亡主因，其中癌症占13 %，而肝癌一直位於惡性腫瘤發生及致死率前五位。從流行病學的研究中，B型肝炎與C型肝炎病毒的感染為導致肝癌的重要危險因子，而亞洲地區為其高盛行率區域，故關於護肝的健康議題更加受到亞洲人的重視。本研究目的以四氯化碳誘導大鼠肝損傷模式，評估由順天生技所提供的中草藥複方(HC)是否具有保肝的功效；HC主要包含維他命C、薑黃及綠茶萃取物這三種已被證實具有抗氧化的功效。由實驗結果顯示，相較於CCl4負對照組，我們發現給予高劑量HC (375 mg/kg bw/day)後，可顯著降低血液中肝臟生化功能指標酵素ALT及AST指數，表示肝損傷獲得改善。而在抗氧化酵素活性測試中，亦可看出HC相較於CCl4負對照組，可提高GPx、GRd、CAT及SOD的活性與GSH含量，表示HC (375 mg/kg bw/day)具有恢復抗氧化系統功能的效果。另外HC亦能顯著降低肝臟中TBARS及TG的含量，顯示HC可以改善脂肪過氧化的現象；組織切片的部分，HC對肝臟空泡化有降低的趨勢，顯示其對肝臟之脂肪變性具有抑制的效果，而高劑量樣品具有降低肝纖維化之潛力。從以上結果顯示中草藥複方HC應對護肝有正面的助益。 2. 肝癌與肝相關疾病為台灣死亡人口中的主要死因，且亞洲人罹患肝炎的比例是世界為最，慢性的B、C型肝炎患者為演變成肝癌的高危險群，此外，在臨床上，治療肝癌的藥物往往具有副作用，目前傳統的癌症治療方法，常引起嚴重的副作用(side effects)，導致病人無法負荷甚至威脅到性命，而依賴醫療方式，癒後病患之五年存活率平均約40%。近年來科學家致力研究癌症的分子及基因調控機制，並積極找尋可減輕癌症症狀或是延緩腫瘤生長速度的療法，現今著重於利用天然物質(natural products)達到癌症預防及促進健康的效果且無副作用之影響，而食用天然食品已為現代人生活保健的趨勢。此外，西方國家亦漸漸對於亞洲傳統中草藥(traditional Chinese medicine)賦予肯定，認為某些癌症發生率較低可能基於亞洲人的飲食因素，故揭開complementary and alternative medicine (CAM)治療功能的潛力。 巴西洋菇發酵產物之乙醇萃出物(ABE)，在體外MTT實驗中，對人類肝癌細胞Hep 3B cells具有毒殺作用，IC50為138 μg/ml。利用MTT實驗推估裸鼠之動物實驗所需劑量分別為60, 120, 600 mg/kg/day，在接種腫瘤前，先給於ABE兩週，觀察ABE是否具有預防肝癌效果。實驗發現ABE對腫瘤的抑制率為42%，對罹患肝癌裸鼠的生存率可提高56%，此外，相對於正對照組，ABE對於裸鼠的的身體負荷較小，體重較正對照組為重且飲食狀況較佳。因此根據活體實驗，ABE似乎具有抗肝癌的潛力。
並列摘要
1. Herbal complex (HC) was offered by Sun-Ten Phytotech. Company. It consists of Vitamin C, green tea, and turmeric which were reported previously with the antioxidant capability. The aim of this study was to evaluate the protective effect of HC against carbon tetrachloride (CCl4)-induced chronic liver damage in rats. Experimental groups were: (1) normal control, (2) Silymarin-positive control, (3) CCl4 -treated group, and (4) HC (4.2-375 mg/kg bw/day) + CCl4 treated groups. After administrated with CCl4 alone on rats for 8 weeks, marked rises of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride, cholesterol, and lipid peroxidation were noted; while the activities of antioxidant enzymes such as glutathione peroxidase (GPx), GRd (glutathione rductase), superoxide dismutase (SOD), and catalase (CAT) were decreased. HC (375 mg/kg bw/day) attenuated successfully all of the negative effects stated above of CCl4 in rats and also elevated the glutathione (GSH) content and inhibited the formation of steatosis as well as fibrosis in the liver as compared with the CCl4 group (p<0.05). These results suggest HC may usefully protect the liver against the oxidative stress induced by CCl4. 2. Over the last decade, the basidiomycete fungus Agaricus blazei Murill has been studied and developed as a novel functional food in some Asian countries after it was reported to be a source of anti-tumor and immuno-active compounds. According to tradition, it also helps against a variety of diseases, including diabetes, hyperlipidemia, atherosclerosis and chronic hepatitis. However, the fruiting body of A. blazei is relative expensive, due to its low yield, and a cheap and stable source of A. blazei fermentation products is desirable to promote in commercial products. In Taiwan, the incidence of liver diseases is high, such as chronic hepatitis, liver cirrhosis and hepatoma which are the leading causes of death in 2006. Furthermore, the effect of the fermentation product of A. blazei cultured in the medium containing soybean and black soybean on liver tumor, we use nude mice with immunodeficiency as a hepatocellular carcinoma (HCC)-induced model. In vitro assay, we found that the ethanolic extract of the fermentation product of A. blazei (ABE) can inhibit Hep 3B cells with IC50, 138 μg/ml. Prior to tumor-inducing, the nude mice received oral administration of ABE in two weeks with preventive goal of food. In vivo model, oral administration of ABE has potential to reduce the tumor volume as compared with negative control and the inhibition rate is 42% on day 51 after tumor-inducing. Moreover, ABE also prolongs the survival time of tumor-bearing mice and the survival rate is 56%. In conclusion, ABE may show the potency to prevent from HCC and is possible to be a candidate for increasing the life span of patients who suffer liver cancer.