Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/66026
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
引用: Bharti, A. C., and B. B. Aggarwal (2002) Nuclear factor-kappa B and cancer: its role in prevention and therapy. Biochem Pharmacol, 64: 883-888. Braudeau, C., D. Bouchet, C. Toquest, L. Tesson, S. Menoret, S. Iyer, C. Loboisse, D. Willis, A. Jarry, R. Buelow, I. Anegon, and C. Chauveau (2003) Generation of the heme oxygenase-1-transgenic rats. Exp Biol Med., 228: 466-471. Chang, C. Y., Z. N. Huang, H. H. Yu, L. H. Chang, S. L. Li, Y. P. Chen, K. Y. Lee, and J. J. Chuu (2008) The adjuvant effects of Antrodia Camphorata extracts combined with anti-tumor agents on multidrug resistant human hepatoma cells. J. Ethnopharmacology, 118(3): 387-395. Chang, J. M., Y. R. Lee, K. M. Hung, S. Y. Liu, M. T. Kuo, W. C. Wen, and Peini Chen (2011) An extract of Antrodia camphorata mycelia attenuates the progression of nephritis in systemic luous erythematosus-prone NZB/W F1 Mice. Evidence-Based Complementary and Alternative Medicine, 10: 1093-1099. Chen, Y. S., J. H. Pan, B. H. Chiang, F. J. Lu, and L. Y. Sheen (2008) Ethanolic extracts of Antrodia cinnamomea mycelia fermented at varied times and scales have differential effects on hepatoma cells and normal primary hepatocytes. J. Food Science, 73(7): H179-H185. Daz, S. K., and D. M. Vasudevan (2007) Alcohol-induced oxidative stress. Life Sci., 81: 177-187. Fernandez-Checa, J. C., A. Colell, and C. Garcia-Ruiz (2002) S- L-methionine and mitochondrial reduced glutathione depletion in alcoholic liver disease. Alcohol, 27: 179-183. Farombi, E. O., S. Shrotriya, H. K. Na, S. H. Kim, and Y.J. Surh (2008) Curcumin attenuates dimethylnitrosamine-indued liver injury in rats through Nrf-2-mediated induction of heme oxygenase-1. Food Chem Toxicol, 46: 1279-1287. Ho, C. M., C. C. Huang, C. J. Huang, J. S. Cheng, I. S. Chen, J. Y. Tsai, B. P. Jiann, P. L. Tseng, S. J. Kuo, and C. R. Jan (2008) Effects of Antrodia camphorata on viability, apoptosis, and [Ca2+]i in PC3 human prostate cancer cells. Chin J Physiol., 51(2): 78-84. Hseu, Y. C., W. C. Chang, Y. T. Hseu, C. Y. Lee, Y. J. Yech, P. C. Chen, J. Y. Chen, and H. L. Yang (2002) Protection of oxidative damage by aqueous extract from Antrodia camphorata mycelia in normal human erythrocytes. Life Sciences, 71(4): 469-482. Hseu, Y. C., S. C. Chen, H. C. Chen, J. W. Liao, and H. L. Yang (2008) Antrodia camphorata inhibits proliferation of human breast cancer cells in vitro and in vivo. Food and Chemical Toxicology, 46(8): 2680-2688. Hsieh, Y. H., F. H. Chu, Y. S. Wang, S. C. Chien, S. T. Chang, J. F. Shaw, C. Y. Chen, W. W. Hsiao, Y. H. Kuo, and S. Y. Wang (2010) Antrocamphin A, an anti-inflammatory principal from the fruiting body of Taiwanofungus camphoratus, and its mechanisms. J. Agric Food Chem., 58(5): 3153-3158. Hsu, Y. L., P. L. Kuo, C. Y. Cho, W. C. Ni, T. F. Tzeng, L. T. Ng, Y. H. Kuo, and C. C. Lin (2007) Antrodia cinnamomea fruiting bodies extract suppresses the invasive potential of human liver cancer cell line PLC/PRF/5 through inhibition of nuclear factor kappa B pathway. Food and Chemical Toxicology, 45(7): 1249-1257. Hu, R., T. O. Khor, G. Shen, W. S. Jeong, V. Hebbar, and Chen C (2006) Cancer chemoprevention of intestinal polyposis in ApcMin/+ mice by sulforaphane, a natural product derived from cruciferous vegetable. Carcinogenesis, 27: 2038-2046. Huang, C. C., H. H. Cheng, J. L. Wang, J. S. Cheng, K. L. Chai, Y. C. Fang, C. C. Kuo, S. T. Chu, C. M. Ho, K. L. Lin, J. Y. Tsai, and C. R. Jan (2009) Effects of Antrodia camphorata extracts on the viability, apoptosis, [Ca2+](i), and MAPKs phosphorylation of OC2 human oral cancer cells. Chinese J. Physiology, 52(3): 128-135. Huang, C. H., Y. Y. Chang, C. W. Liu, W. Y. Kang, Y. L. Lin, H. C. Chang, and Y. C. Chen (2010) Fruiting Body of Niuchangchih (Antrodia camphorata) Protects Livers against Chronic Alcohol Consumption Damage. J. Agric. and Food Chem., 58(6): 3859-3866. Kuo, P. L., Y. L. Hsu, C. Y. Cho, L. T. Ng, Y. H. Kuo, and C. C. Lin (2006) Apoptotic effects of Antrodia cinnamomea fruiting bodies extract are mediated through calcium and calpain-dependent pathways in Hep3B cells. Food and Chemical Toxicology, 44(8): 1316-1326. Lee, T. H. (2007) A new cytotoxic agent from solid-state fermented mycelium of Antrodia camphorata (2007) Planta Medica, 73(14): 1522-1522. Lien, H. M., H. W. Lin, Y. J. Wang, L. C. Chen, D. Y. Yang, Y. Y. Lai, and Y. S. Ho (2009) Inhibition of anchorage-independent proliferation and G0/G1 cell-cycle regulation in human colorectal carcinoma cells by 4,7-Dimethoxy-5-methyl-l,3-benzodioxole isolated from the fruiting body of Antrodia camphorate. Evid Based Complement Alternat Med., 3253-3265. Lin, W. C., S. C. Kuo, W. L. Lin, H. L. Fang, and B. C. Wang (2006) Filtrate of fermented mycelia from Antrodia camphorata reduces liver fibrosis induced by carbon tetrachloride in rats. World Journal of Gastroenterology, 12(15): 2369-2374. Liu, J. J., T. S. Huang, M. L. Hsu, C. C. Chen, W. S. Lin, F. J. Lu, and W. H. Chang (2004) Antitumor effects of the partially purified polysaccharides from Antrodia camphorata and the mechanism of its action. Toxicology and Applied Pharmacology, 201(2): 186-193. Liu, K. J., S. J. Leu, C. H. Su, B. L. Chiang, Y. L. Chen, and Y. L. Lee (2010) Administration of polysaccharides from Antrodia camphorata modulates dendritic cell function and alleviates allergen-induced T helper type 2 responses in a mouse model of asthma. Immunology, 129(3): 351-362. Lu, M. C., Y. C. Du, J. J. Chuu, S. L. Hwang, P. C. Hsieh, C. S. Hung, F. R. Chang, and Y. C. Wu (2009) Active extracts of wild fruiting bodies of Antrodia camphorata (EEAC) induce leukemia HL60 cells apoptosis partially through histone hypoacetylation and synergistically promote anticancer effect of trichostatin A. Archives of Toxicology, 83(2): 121-129. Martin, A. R., I. Villegas, M. Sanchez-Hidalgo, and C. A. Lastra (2006) The effects of resveratrol, a phytoalexin derived from red wines, on chronic inflammation induced in an experimentally induced colitis model. Br J. Pharmacol, 147: 873-885. Ogborne, R. M., S. A. Rushworth, and M. A. O'Connell (2008) Epigallocatechin activates haem oxygenase-1 expression via protein kinase C and Nrf-2. Biochem Biophys Res Commun, 373: 584-588. Peng, C. C., K. C. Chen, R. Y. Peng, C. C. Chyau, C. H. Su, and H. M. Hsieh-Li (2007) Antrodia camphorata extract induces replicative senescence in superficial TCC, and inhibits the absolute migration capability in invasive bladder carcinoma cells. J. Ethnopharmacology, 109(1): 93-103. Peng, C. C., K. C. Chen, R. Y. Peng, C. H. Su, and H. M. Hsieh-Li (2006) Human urinary bladder cancer T24 cells are susceptible to the Antrodia camphorata extracts. Cancer Letters, 243(1): 109-119. Rao, Y. K., A. T. H. Wu, M. Geethangili, M. T. Huang, W. J. Chao, C. H. Wu, W. P. Deng, C. T. Yeh, and Y. M. Tzeng (2011) Identification of antrocin from Antrodia camphorata as a selective and novel class of small molecule inhibitor of Akt/mTOR signaling in metastatic breast cancer MDA-MB-231 Cells. Chemical Research in Toxicology, 24(2): 238-245. Kumar, K.J. S, F. H. Chu, H. W. Hsieh, J. W. Liao, W. H. Li, J. Lin, J. F. Shaw, and S. Y. Wang (2011) Antroquinonol from ethanolic extract of mycelium of Antrodia cinnamomea protects hepatic cells from ethanol-induced oxidative stress through Nrf-2 activation. J. Ethnopharmacology, 10: 1016-1027. Shen, Y. C., C. J. Chou, Y. H. Wang, C. F. Chen, Y. C. Chou, and M. K. Lu (2004) Anti-inflammatory activity of the extracts from mycelia of Antrodia camphorata cultured with water-soluble fractions from five different Cinnamomum species. Fems Microbiology Letters, 231(1): 137-143. Shishodia, S., H. M. Amin, R. Lai, and B. B. Aggarwal (2005) Curcumin (diferuloylmethane) inhibits constitutive NF-kappaB activation, induces G1/S arrest, suppresses proliferation, and induces apoptosis in mantle cell lymphoma. Biochem Pharmacol, 70: 700-713. Song, T. Y., S. L. Hsu, C. T. Yeh, and G. C. Yen (2005) Mycelia from Antrodia camphorata in submerged culture induce apoptosis of human hepatoma HepG2 cells possibly through regulation of Fas pathway. J. Agric. and Food Chem., 53(14): 5559-5564. Song, T. Y., and G. C. Yen (2003) Protective effects of fermented filtrate from Antrodia camphorata in submerged culture against CCl4-induced hepatic toxicity in rats. J. Agric. and Food Chem., 51(6): 1571-1577. Sporn, M. B., N. M. Dunlop, D. L. Newton, and J. M. Smith (1976) Prevention of chemical carcinogenesis by vitamin A and its synthetic analogs (retinoids). Fed Proc., 35: 1332-1338. Surh, Y. J. (2003) Cancer chemoprevention with dietary phytochemicals. Nat Rev Cancer, 3: 768-780. Yang, F., W. Villiers, C. J. McClain, and G. W. Varilek (1998) Green tea polyphenols block endotoxin-induced tumor necrosis factor-production and lethality in a murine model. J. Nutr., 128: 2334-2340. Yang, H. L., C. S. Chen, W. H. Chang, F. J. Lu, Y. C. Lai, C. C. Chen, T. H. Hseu, C. T. Kuo, and Y. C. Hseu (2006) Growth inhibition and induction of apoptosis in MCF-7 breast cancer cells by Antrodia camphorata. Cancer Letters, 231(2): 215-227. Yang, S. S., G. J. Wang, S. Y. Wang, Y. Y. Lin, Y. H. Kuo, and T. H. Lee (2009) New constituents with iNOS inhibitory activity from mycelium of Antrodia camphorata. Planta Medica, 75(5): 512-516. Yeh, C. T., C. J. Yao, J. L. Yan, S. E. Chuang, L. M. Lee, C. M. Chen, C. F. Yeh, C. H. Li, and G. M. Lai (2011) Apoptotic cell death and inhibition of Wnt/beta-Catenin signaling pathway in human colon cancer cells by an active fraction (HS7) from Taiwanofungus camphoratus. Evid Based Complement Alternat Med., 2011: 750230. Yeh, C. T., Y. K. Rao, C. J. Yao, C. F. Yeh, C. H. Li, S. E. Chuang, J. H. T. Luong, G. M. Lai, and Y. M. Tzeng (2009) Cytotoxic triterpenes from Antrodia camphorata and their mode of action in HT-29 human colon cancer cells. Cancer Letters, 285(1): 73-79.
摘要: 
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.
URI: http://hdl.handle.net/11455/66026
其他識別: U0005-2308201123384200
Appears in Collections:森林學系

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