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標題: 固態培養牛樟芝抗發炎及抗氧化活性
Anti-inflammatory and Anti-oxidative Activities of Antrodia cinnamomea Mycelium Producing by Solid-state Culture
作者: 謝翰文
Hsieh, Han-Wen
關鍵字: Antrodia cinnamomea;牛樟芝;anti-inflammatory;hepaprotective;4-acetyl-antroquinonol B;抗發炎;肝臟保護;4-Acetyl-antroquinonol B
出版社: 森林學系所
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牛樟芝(Antrodia cinnamomea)是一種傳統的藥用真菌類,對腹瀉、腹痛、高血壓、糖尿病、尿蛋白、肝硬化、肝癌等症狀或疾病具有非常良好的療效。因此本實驗以固態培養之牛樟芝菌絲體為主題,利用管柱層析、薄層層析、高效能液相層析儀及核磁共振質譜儀等對其主要成分做鑑定和定量。由分析結果整理,固態培養牛樟芝之主要成分為Cytosine (0.21 μg/mg)、Uracil (0.15 μg/mg)、Cytidine (0.76 μg/mg)、Uridine (1.24 μg/mg)、Adenine (0.06 μg/mg)、Inosine (1.61 μg/mg)、Guanosine (0.71 μg/mg)、Adenosine (0.61 μg/mg)、Deoxyadenosine (1.03 μg/mg)、Dehydroeburicoic acid (0.95 μg/mg)、Dehydrosulphurenic acid (0.91 μg/mg)、3-Isobutyl-4-furan-2,5-dione (1.31 μg/mg)、Antroquinonol (1.78 μg/mg) 和4-Acetyl-antroquinonol B (1.48 μg/mg)等。
在生物活性的部分,本論文則分為兩個主題,第一個主題是牛樟芝的抗發炎活性,首先我們以牛樟芝菌絲體之粗萃物對LPS誘導體內發炎之ICR小鼠做體內抗發炎活性的分析,可以明顯觀察牛樟芝菌絲體抽出物可以有效的抑制由LPS誘導發炎所活化之NF-κB路徑及其下游之iNOS、COX-2蛋白的表現。接著續以生物活性為導向配合小鼠巨噬細胞RAW264.7做抗發炎反應分析,我們從牛樟芝菌絲體萃取物中獲得了能夠有效抑制發炎反應的化合物4-Acetyl-antroquinonol B,並在體外試驗證實,4-Acetyl-antroquinonol B能有效透過抑制NF-κB路徑的表現,進而抑制下游iNOS蛋白表現合一氧化氮自由基的產生。因此我們得知無論在體內試驗還是體外試驗,牛樟芝菌絲體都能夠提供優良的抗發炎能力。
另一方面本論文透過了動物試驗及細胞試驗證實固態培養之牛樟芝菌絲體粗萃物,具有保護動物肝臟因酒精誘導產生病變之活性。在動物試驗中,本研究使用乙醇 (80 % v/v) 來誘導ICR公鼠之肝臟病變。而由一些與乙醇誘導肝臟病變相關之細胞激素,包括MDA、ALT和AST等的表現得知,牛樟芝菌絲體萃取物可以顯著地對肝臟細胞產生保護作用並呈現劑量相關性。此外,牛樟芝菌絲體萃取物可增加肝臟細胞在酒精傷害後穀胱甘

Antrodia cinnamomea is endemic medicine fungus of Taiwan. It has been claimed for having significant effects on diarrhea, abdominal pain, high blood pressure, diabetes, urinates the protein, liver cirrhosis and liver cancer. In this study, we choose Antrodia cinnamomea mycelium producing by solid-state culture was selected as the target for investigate. First, the composition of extract prepared from mycelium was analyzed by HPLC. The results revealed that the principal components of mycelium is cytosine (0.21 μg/mg), uracil (0.15 μg/mg), cytidine (0.76 μg/mg), uridine (1.24 μg/mg), adenine (0.06 μg/mg), inosine (1.61 μg/mg), guanosine (0.71 μg/mg), adenosine (0.61 μg/mg), deoxyadenosine (1.03 μg/mg), dehydroeburicoic acid (0.95 μg/mg), dehydrosulphurenic acid (0.91 μg/mg), 3-isobutyl-4-furan-2,5-dione (1.31 μg/mg), antroquinonol (1.78 μg/mg) and 4-acetyl-antroquinonol B (1.48 μg/mg).
As regard to the bioactivity study, there are two subjects, anti-inflammatory and anti-oxidative activities, were evaluated in this study. The model of LPS-induced acute inflammation in ICR mice was conducted to the anti-inflammatory activity of Antrodia cinnamomea (EMAC) in vivo. According to the results obtained from protein expression assay, EMAC presented a significant inhibitory activity for NF-κB, as well as its downstream proteins, iNOS and COX-2. Furthermore, an anti-inflammation compound, 4-acetyl-antroquinonol B was obtained from EMAC by using LPS-induced inflammation on macrophage RAW264.7 cell model. 4-acetyl-antroquinonol B revealed the strong activity to inhibit NF-κB and iNOS expression, by this reducing NO production.
On the other hand, the heapprotective effect of EMAC on mice liver damaged by oxidative stress was investigated in this study. The animal model of alcohol-induced liver damage in ICR mice was used to evaluate the activity of EMAC. When the mice pretreated EMAC, the liver oxidative damaged related cytokines, including MDA, ALT and AST were significantly reduced with dose-dependent manner comparing without EMAC pretreated mice. Besides, the level of glutathione (GSH) was reduced after the mice received EMAC. Moreover, the EMAC treated groups could increase the HO-1 and Nrf-2 proteins expression. The anti-inflammation compound, 4-acetyl-antroquinonol B, also revealed the hepaprotective effect on HepG2 cell after alcohol-induced damage.
其他識別: U0005-2308201123384200
Appears in Collections:森林學系

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