| 摘要: | 中文摘要
本研究目的首先以高效液相層析質譜儀分析法，建立梅里爾針層孔菌的乙醇抽出物之指紋圖譜。其次比較其不同溶劑分層的萃取物之體外抗氧化活性，最後探討梅里爾針層孔菌的乙醇抽出物對肝臟保護、鎮痛、抗發炎作用及抗肝腫瘤活性。
本研究的研究方法如下:在體外抗氧化方面有ABTS自由基清除測定、測定總酚類含量、FRAP測試、快速DPPH呈色反應、DPPH清除自由基活性和還原力等項目。在急性肝炎方面，檢驗血清中GOT、GPT、肝臟組織SOD、GPX、catalase數據及肝臟組織切片H.E.染色。在鎮痛與抗發炎方面以醋酸誘導扭體試驗和福馬林注射足蹠之舔足時間測試及λ-角叉菜膠誘導足蹠浮腫測試。在抗肝腫瘤活性方面，檢驗血清中GOT、GPT、γ-GT和肝臟組織切片中H.E.病理變化與γ-GT染色活性分析及NK-cell活性分析測試。
本研究結果，在高效液相層析質譜儀分析法中，hispolon與梅里爾針層孔菌的乙醇抽出物顯示出有相似的波峰與相同的4.7分鐘的滯留時間，均獲得m/z 219與m/z 135有較大的裂片。本研究在體外抗氧化方面，以EtOAC及n-BuOH分層的抗氧化活性較強。在急性肝炎方面，血清中GOT、GPT、肝臟組織SOD、GPX、catalase數據，依給藥劑量的增加而減少數值，及肝臟組織切片H.E.染色方面，給藥組傷害有減輕的趨勢。在鎮痛與抗發炎方面，依給藥劑量的增加對小鼠醋酸扭體反應次數有顯著抑制作用，對福馬林誘導的後期疼痛有明顯的抑制效果，在抗發炎方面，會抑制λ-角叉菜膠誘導的足部浮腫。在抗肝腫瘤活性方面，檢驗血清中GOT、GPT、γ-GT依給藥劑量的增加而降低其指數，肝臟組織切片中H.E.與γ-GT染色結果於給藥組其傷害有減輕的趨勢，而NK-cell活性測試結果呈現給藥組有提升NK細胞撲殺YAC-1腫瘤細胞株的趨勢。
綜合上述研究結果顯示，梅里爾針層孔菌乙醇抽出物含有活性成份hispolon，其EtOAc及n-BuOH分層具有抗氧化活性。梅里爾針層孔菌的乙醇抽出物對四氯化碳誘發的急性肝損傷有保護作用，並有鎮痛、抗發炎、抗肝腫瘤及增強細胞免疫力的活性，其活性成分為hispolon。
Abstract
The main aims of this study included establishing the ion fingerprinting pattern of Phellinus merrillii extracts by high-performance liquid chromatography tandem mass (LC/MS/MS) analysis, and evaluation of potential antioxidant activity of fraction components in P. merrillii. The effects of P. merrillii ethanol extracts on antioxidation, analgesia, anti-inflammation and hepatoprotection by carbon tetrachloride-induced hepatotoxicity were evaluated. Further assessment of the anti-carcinogenic activity and the mechanisms of its chemopreventive action were also performed.
The antioxidative assays performed included: ABTS free radical scavenging assay, determination of total phenolics contents (TPC), ferric reducing antioxidant power assay (FRAP), rapid screening of antioxidant by dot-blot DPPH (1,1-diphenyl-2-picrylhydrazyl) staining, DPPH radical-scavenging activities and reducing power measurement. In the hepatoprotective studies, serum levels of GOT, GPT, the liver SOD, and GPX, the catalase activities and the liver tissue examinations were determined. The full time lick induction test and the γ-carrageenin foot edema induction test were also performed. In the aspect of anti-liver tumor capacity, serum GOT, GPT, γ-GT measurement and liver tissue examination, together with NK-cell activity were determined.
The results showed that P. merrillii extracts did contain the active ingredient hispolon. Significant in vitro oxidative resistance phenomena by EtOAC and n-BuOH fractions were observed. Liver functional, inflammation and morphological studies showed reduction of liver damage by administration of P. merrillii extracts. Accordingly, analgesia and foot edema reduction together with anti-tumor activity were also significantly observed. In comparison, the NK-cell activity was increased.
In conclusion, P. merrillii extracts contained active compound, hispolon, and has the strong anti-oxidation, liver protection, analgesia, anti-inflammation and anti-hepatoma activities. |
