臺灣博碩士論文加值系統

本篇研究是在探討椴木栽培的牛樟芝(Taiwanofungus camphoratus)子實體超臨界萃取物(TCSE)能否改善四氯化碳引起的大鼠肝臟纖維化病症。大鼠每100 g體重經口給予0.2毫升的四氯化碳(20%)，每週兩次，持續八週。每天給予TCSE，八週後犧牲大鼠並採取檢體。第八週的alanine transaminase、aspartate transaminase均有顯著下降，白蛋白則是有顯著回升。肝臟檢體的膠原蛋白含量以及切片檢查均顯示TCSE能顯著抑制膠原蛋白的產生。肝臟檢體的Glutathione顯著回升與Malondialdehyde顯著下降顯示TCSE能顯著提升抗氧化能力。免疫組織染色的部份顯示TCSE能有效降低alpha smooth muscle actin (α-SMA)的產生。RT-PCR的數據顯示TCSE能顯著抑制Col1α1與α-SMA兩個基因的表現。
依據以上數據，推論TCSE抑制四氯化碳誘導的肝纖維化，可能是經由抑制肝星狀細胞(hepatic stellate cell；HSC)的活化來達到保肝作用。

This article investigated whether Taiwanofungus camphoratus carpophore''s supercritical extract(TCSE) can improve liver fibrosis on rats induced by carbon tetrachloride. Rats got 0.2 mL carbon tetrachloride (20%) per 100g body weight orally, twice a week, for eight weeks. Rats treated with TCSE, and were sacrificed. The value of aspartatetransaminase and alanine transaminase on the eighth week decreased significantly, and albumin increased. Collagen quantity of liver samples and slides'' check showed that TCSE can inhibit production of collagen. Glutathione increasing significantly and malondialdehyde decreasing significantly showed that TCSE can promote anti-oxidative activity.Tissue immunochromatography slides showed that TCSE can reduce the production of alpha smooth muscle actin (α-SMA).TCSE can also inhibit mRNA of Col1α1 and α-SMA on RT-PCR Data.
According to above data, we can infer that TCSE can inhibit liver fibrosis process induced by carbon tetrachloride and have liver protection, probably through inhibiting activity of hepatic stellate cell(HSC).

目錄.....................................................V
圖表目錄...............................................VII
中文摘要.................................................IX
英文摘要.................................................X
第一章 緒論...............................................1
一、 研究背景與目的..................................1
二、 牛樟芝文獻考察..................................2
(一) 牛樟芝型態......................................3
(二) 牛樟芝段木栽培..................................4
(三) 牛樟芝化學成分..................................5
(四) 牛樟芝藥理研究..................................5
(五) 純化合物的藥理活性..............................8
三、 肝臟纖維化病理機制.............................16
(一) 肝臟纖維化病理成因.............................16
(二) 四氯化碳動物病理模式...........................18
(三) 抗氧化酵素 ....................................20
第二章 材料與方法........................................22
第三章 結果..............................................30
第四章 討論..............................................34
第五章 參考文獻..........................................53

1.Wen CL, Chang CC, Huang SS, Kuo CL, Hsu SL, Deng JS, Huang GJ. Anti-inflammatory effects of methanol extract of Antrodia cinnamomea mycelia both in vitro and in vivo. J Ethnopharmacol. 2011;137(1):575-84.
2.Lin YW, Chiang BH. 4-acetylantroquinonol B isolated from Antrodia cinnamomea arrests proliferation of human hepatocellular carcinoma HepG2 cell by affecting p53, p21 and p27 levels. J Agric Food Chem. 2011;59(16):8625-31.
3.Weber LW, Boll M, Stampfl A. Hepatotoxicity and mechanism of action of haloalkanes: carbon tetrachloride as a toxicological model. Crit Rev Toxicol. 2003;33(2):105-36.
4.Wu SH, Yu ZH, Dai YC, Chen CT, Su CH, Chen LC, Hsu WC, Hwang GY. Taiwanofungus, a polypore new genus. Fung. Sci. 2004;19:109-116.
5.Geethangili M, Tzeng YM. Review of Pharmacological Effects of Antrodia camphorata and its Bioactive Compounds. Evid Based Complement Alternat Med. 2011;212641.
6.Chang TT, Chou WN. Antrodia cinnamomea sp. nov. on Cinnamomum kanehirai in Taiwan. Mycol. Res. 1995;99:756-758.
7.Hsu YL, Kuo YC, Kuo PL, Ng LT, Kuo YH, Lin CC. Apoptotic effects of extract from Antrodia camphorate fruiting bodies in human hepatocellular carcinoma cell lines. Cancer Letters. 2005;221(1):77–89.
8.Kuo PL, Hsu YL, Cho CY, Ng LT, Kuo YH, Lin CC. Apoptotic effects of Antrodia cinnamomea fruiting bodies extract are mediated through calcium and calpaindependent pathways in Hep 3B cells. Food and Chemical Toxicology. 2006;44(8):1316–1326.
9.Rao YK, Fang SH, Tzeng YM. Evaluation of the anti-inflammatory and anti-proliferation tumoral cells activities of Antrodia camphorata, Cordyceps sinensis, and Cinnamomum osmophloeum bark extracts. Journal of Ethnopharmacology. 2007;114(1):78–85.
10.Shen YC, Chou CJ, Wang YH, Chen CF, Chou YC, Lu MK. Anti-inflammatory activity of the extracts from mycelia of Antrodia camphorata cultured with watersoluble fractions from five different Cinnamomum species. FEMSMicrobiology Letters. 2004;231(1):137–143.
11.Hseu YC, Wu FY, Wu JJ, Chen JY, Chang WH, Lu FJ, Lai YC, Yang HL. Anti-inflammatory potential of Antrodia camphorata through inhibition of iNOS, COX-2 and cytokines via the NF-κB pathway. International Immunopharmacology. 2005;5(13-14):1914–1925.
12.Rao YK, Fang SH, Tzeng YM. Evaluation of the anti-inflammatory and anti-proliferation tumoral cells activities of Antrodia camphorata, Cordyceps sinensis, and Cinnamomum osmophloeum bark extracts. Journal of Ethnopharmacology. 2007;114(1):78–85.
13.Chen JC, Chen CN, Sheu SJ et al. Liver-caring medicine containing Antrodia camphorate. 2003;US 0113297(003):6-19.
14.Huang RL, Huang Q, Chen CF, Chang TT, hou CJ. Anti-viral effects of active compounds from Antrodia amphorata on wild-type and lamivudine-resistant mutant BV. Chinese Pharmaceutical Journal. 2003;55(5):371-379.
15.Shu CH, Lung MY. Effect of culture pH on the antioxidant properties of Antrodia camphorata in submerged culture. Journal of the Chinese Institute of Chemical Engineers. 2008;39(1):1-8.
16.Song TY, Yen GC. Antioxidant properties of Antrodia amphorata in submerged culture. Journal of Agricultural and Food Chemistry. 2002;50(11):3322-3327.
17.Hseu YC, Chen SC, Yech YJ, Wang L, Yang HL. Antioxidant activity of Antrodia camphorata on free radical-induced endothelial cell damage. Journal of Ethnopharmacology. 2008;118(2):237-245.
18.Dai YY, Chuang CH, Tsai CC et al. The protection of Anthrodia camphorata against acute hepatotoxicity of alcohol in rats. Journal of Food and Drug Analysis. 2003;11:177-185.
19.Hsiao G, Shen MY, Lin KH, Lan MH, Wu LY, Chou DS, Lin CH, Su CH, Sheu JR. Antioxidative and hepatoprotective effects of Antrodia camphorata extract. Journal of Agricultural and Food Chemistry. 2003;51(11):3302-3308.
20.Song TY, Yen GC. Protective effects of fermented iltrate from Antrodia camphorata in submerged culture gainst CCl4-induced hepatic toxicity in rats. Journal of gricultural and Food Chemistry. 2003;51(6): 1571– 577.
21.Huang JS, Chang HC, Li EIC, Huang TM, Su YH, and Wang KC. Enhancement of hepatoprotective efficacy of Antrodia camphorata by Chinese tradition medicine. Journal of Gastroenterology and Hepatology. 2006;21(supplement 2): A234.
22.Lin WC, Kuo SC, Lin WL, Fang HL, Wang BC. Filtrate of fermented mycelia from Antrodia camphorata reduces liver fibrosis induced by carbon tetrachloride in rats. World Journal of Gastroenterology. 2006;12(15): 2369–2374.
23.Bataller R. Brenner DA. Liver fibrosis. J Clin Invest. 2005;115(2):209-18.
24.Arthur MJ. Fibrogenesis II. Metalloproteinases and their inhibitors in liver fibrosis. Am J Physiol Gastrointest Liver Physiol. 2000;279(2):G245-9.
25.Gabele E, Brenner DA, Rippe RA. Liver fibrosis: signals leading to the amplification of the fibrogenic hepatic stellate cell. Front Biosci. 2003;8:d69-77.
26.Marra F. Hepatic stellate cells and the regulation of liver inflammation. J Hepatol. 1999;31(6):1120-30.
27.Kmieć Z. Cooperation of liver cells in health and disease. Adv Anat Embryol Cell Biol. 2001;161:III-XIII, 1-151.
28.Higuchi H, Gores GJ. Mechanisms of liver injury: an overview. Curr Mol Med. 2003;3(6):483-90.
29.Canbay A, Friedman S, Gores GJ. Apoptosis: the nexus of liver injury and fibrosis. Hepatology. 2004;39(2):273-8.
30.Casini A, Ceni E, Salzano R, Biondi P, Parola M, Galli A, Foschi M, Caligiuri A, Pinzani M, Surrenti C. Neutrophil-derived superoxide anion induces lipid peroxidation and stimulates collagen synthesis in human hepatic stellate cells: role of nitric oxide. Hepatology. 1997;25(2):361-7.
31.Maher JJ. Interactions between hepatic stellate cells and the immune system. Semin Liver Dis. 2001;21(3):417-26.
32.Naito M, Hasegawa G, Ebe Y, Yamamoto T. Differentiation and function of Kupffer cells. Med Electron Microsc. 2004;37(1):16-28.
33.Neuman Re, Logan Ma. The determination of hydroxyproline. J Biol Chem. 1950;184(1):299-306.
34.Dr. Emilio Orfei. REVIEW OF PATHOLOGY OF THE LIVER. Loyola University of Chicago Stritch School of Medicine Department of Pathology
35.Estrela JM, Ortega A, Obrador E. Glutathione in cancer biology and therapy. Crit Rev Clin Lab Sci. 2006;43(2):143-81.
36.De Mattia G, Bravi MC, Laurenti O, Cassone-Faldetta M, Armiento A, Ferri C, Balsano F. Influence of reduced glutathione infusion on glucose metabolism in patients with non-insulin-dependent diabetes mellitus. Metabolism. 1998;47(8):993-7.
37.Palamara AT, Perno CF, Aquaro S, Bue MC, Dini L, Garaci E. Glutathione inhibits HIV replication by acting at late stages of the virus life cycle. AIDS Res Hum Retroviruses. 1996;12(16):1537-41.
38.Bounous G, Batist G, Gold P. Immunoenhancing property of dietary whey protein in mice: role of glutathione. Clin Invest Med. 1989;12(3):154-61.
39.Schulz JB, Lindenau J, Seyfried J, Dichgans J. Glutathione, oxidative stress and neurodegeneration. Eur J Biochem. 2000;267(16):4904-11.
40.Powers SK, Jackson MJ. Exercise-induced oxidative stress: cellular mechanisms and impact on muscle force production. Physiol Rev. 2008 ;88(4):1243-76.
41.Aebi H. Catalase in vitro. Methods Enzymol. 1984;105:121-6.
42.Yamada Y, Fausto N. Deficient liver regeneration after carbon tetrachloride injury in mice lacking type 1 but not type 2 tumor necrosis factor receptor. Am J Pathol. 1998;152(6):1577-89.
43.Perez Tamayo R. Is cirrhosis of the liver experimentally produced by CCl4 an adequate model of human cirrhosis? Hepatology. 1983;3(1):112-20.
44.Chen CJ, Su CH, Lan MH. Study on solid cultivation and bioactivity of Antrodia camphorate. Fungal Sciences. 2010;16:65-72.
45.Chen CH, Yang SW, Shen YC. New steroid acids from Antrodia cinnamomea, a fungal parasite of Cinnamomum micranthum. Journal of Natural Products. 1995;58(11):1655-1661.
46.Yeh CT, Rao YK, Yeh CF et al. Cytotoxic triterpenes from Antrodia camphorata and their mode of action in HT- 29 human colon cancer cells. Cancer Letters. 2009;285(1):73-79.
47.Geethangili M, Fang SH et al. Inhibitory effect of Antrodia camphorate constituents on the Helicobacter pylori-associated gastric inflammation. Food Chemistry. 2010;119(1):149-153.
48.Chen JJ, Lin WJ et al. Antiinflammatory benzenoids from Antrodia camphorate. Journal of Natural Products. 2007;70(6):989-992.
49.Chen YC, Chen CF et al. Evaluation of the immuno-modulating activity of some active principles isolated from the fruiting bodies of Antrodia camphorate. Chinese Pharmaceutical Journal. 2003;55(5):313-318.
50.Shen YC, Wang YH, Chou YC, Chen CF, Lin LC, Chang TT, Tien JH, Chou CJ. Evaluation of the anti-inflammatory activity of zhankuic acids isolated from the fruiting bodies of Antrodia camphorate. Planta Medica. 2004;70(4):310-314.
51.Chen CC, Shiao YJ, Lin RD et al. Neuroprotective diterpenes from the fruiting body of Antrodia camphorate. Journal of Natural Products. 2006;69(4):689-691.
52.Huang KF, Huang WM, Chiang HC.Phenyl compounds from Antrodia cinnamomea. Chinese Pharmaceutical Journal. 2001;53(6):327-331.
53.Shen CC, Yang HC, Huang RL et al. Anti-HBV principle from the culture broth of Antrodia camphorata (strain # CCRC-35396). The Journal of Chinese Medicine. 2005;16:57-61.
54.Chen CC, Chyau CC, Hseu TH. Production of a COX-2 inhibitor, 2,4,5-trimethoxybenzaldehyde, with submerged cultured Antrodia camphorate. Letters in Applied Microbiology. 2007;44(4):387-392.
55.Chien SC, Chen ML et al. Anti-inflammatory activities of new succinic and maleic derivatives from the fruiting body of Antrodia camphorate. Journal of Agricultural and Food Chemistry. 2008;56(16):7017-7022.
56.Wang GJ, Tseng HW et al. The vasorelaxation of Antrodia camphorate mycelia: involvement of endothelial Ca2+-NO-cGMP pathway. Life Sciences. 2003;73(21):2769-2783.
57.Lee IH, Huang RL et al. Antrodia camphorata polysaccharides exhibit anti-hepatitis B virus effects. FEMS Microbiology Letters. 2002;209(1):63-67.
58.Schepetkin IA, Quinn MT. Botanical polysaccharides: macrophage immunomodulation and therapeutic potential. International Immunopharmacology. 2006;6(3):317-333.
59.Liu JJ, Huanf TS et al. Antitumor effects of the partially purified polysaccharides from Antrodia camphorata and the mechanism of its action. Toxicology and Applied Pharmacology. 2004;201(2):186-193.
60.Cheng JJ, Huanf NK et al. Study for anti-angiogenic activities of polysaccharides isolated from Antrodia cinnamomea in endothelial cells. Life Sciences. 2005;76(26):3029-3042.
61.Chen CC, Liu YW, Ker YB et al. Chemical characterization and anti-inflammatory effect of polysaccharides fractionated from submerge-cultured Antrodia camphorate mycelia. Journal of Agricultural and Food Chemistry. 2007;55(13):5007-5012.
62.Chen YJ, Cheng PC, Lin CN et al. Polysaccharides from Antrodia camphorata mycelia extracts possess immunomodulatory activity and inhibits infection of Schistosoma mansoni. International Immunopharmacology. 2008;8(3):458-467.
63.Tsai MC, Song TY, Shih PH et al. Antioxidant properties of water-soluble polysaccharides from Antrodia cinnamomea in submerged culture. Food Chemistry. 2007;104(3):1115-1122.
64.Cheng JJ, Huang NK, Lur HS et al. Characterization and biological functions of sulfated polysaccharides from sulfated-salt treatment of Antrodia cinnamomea. Process Biochemistry. 2009;44(4):453-459.
65.Huang RL, Huang Q, Chen CF et al. Anti-viral effects of active compounds from Antrodia camphorata on wild-type and lamivudine-resistant mutant HBV. Chinese Pharmaceutical Journal. 2003;55(5):371-379.
66.Lee TH, Lee CK, Tsou WL et al. A new cytotoxic agent from solid-state fermented mycelium of Antrodia camphorate. Planta Medica. 2007;73(13):1412-1415.
67.Chang JM, Lee YR, Hung LM et al. An extract of Antrodia camphorata mycelia attenuates the progression of nephritis in systemic lupus erythematosus-prone NZB/W F1 mice. Evidence-Based Complementary and Alternative Medicine. 2011:465894.
68.Liu SY, Kuo MT, Wen WC. Inhibition of hepatitis B virus by cyclohexenone compounds from Antrodia camphorate. 2009:US US 7411003.
69.Yang SS, Wang GJ,Wang SY et al. New constituents with iNOS inhibitory activity from mycelium of Antrodia camphorate. Planta Medica. 2009;75(5):512-516.
70.Lien HM, Lin HW, Wang YJ et al. Inhibition of anchorage-independent proliferation and G0/G1 cellcycle regulation in human colorectal carcinoma cells by 4,7-dimethoxy-5-methyl-l,3-benzodioxole isolated from the fruiting body of Antrodia camphorate. Evidence-Based Complementary and Alternative Medicine. 2011:984027.
71.Chiang CH, Wu DP, Cherng IW et al.A sesquiterpene lactone, phenyl and biphenyl compounds from Antrodia cinnamomea. Phytochemistry. 1995;39(3):613-616.
72.Shen YC, Yang SW, Lin CS et al. Zhankuic acid F: a new metabolite from 14 Evidence-Based Complementary and Alternative Medicine a formosan fungus Antrodia cinnamomea. Planta Medica. 1997;63(1):86-88.
73.Cherng IH, Wu DP, Chiang HC. Triterpenoids from Antrodia cinnamomea. Phytochemistry. 1996;41(1):263-267.
74.Chen CH, Yang SW, Shen YC. New steroid acids from Antrodia cinnamomea, a fungal parasite of Cinnamomum micranthum. Journal of Natural Products. 1995;58(11):1655-1661.
75.Shen CC, Kuo YC, Huang RL et al. New ergostane and lanostane from Antrodia camphorate. The Journal of Chinese Medicine. 2003;14:247-58.
76.Male KB, Rao YK, Tzeng YM et al. Probing inhibitory effects of Antrodia camphorata isolates using insect cell-based impedance spectroscopy: inhibition vs chemical structure. Chemical Research in Toxicology. 2008;21(11):2127-2133.
77.Cherng IH, Chiang HC, Cheng MC et al. Three new triterpenoids from Antrodia cinnamomea. Journal of Natural Products. 1995;58(3):365-371.
78.Chen CC, Shiao YJ, Lin RD et al. Neuroprotective diterpenes from the fruiting body of Antrodia camphorate. Journal of Natural Products. 2006;69(4):689-691.
79.Wu MD, Cheng MJ, Wang BC et al. Chemical constituents from the mycelia of Antrodia cinnamomea. Journal of the Chilean Chemical Society. 2007;52(4):1338-1340.
80.Wu DP, Chiang CH. Constituents of Antrodia cinnamomea. Journal of the Chinese Chemical Society. 1995;42:797-800.
81.Hattori M, Sheu CC. Compounds from Antrodia camphorate having anti-inflammatory and anti-tumor activity. 2006:US 7109232.
82.Wu MD, Cheng MJ, Wang BC et al. Maleimide and maleic anhydride derivatives from the mycelia of Antrodia cinnamomea and their nitric oxide inhibitory activities in macrophages. Journal of Natural Products. 2008;71(7):1258-1261.
83.Lu MK, Cheng JJ, Lai WL et al. Adenosine as an active component of Antrodia cinnamomea that prevents rat PC12 cells from serum deprivation-induced apoptosis through the activation of adenosine A2A receptors. Life Sciences. 2006;79(3):252-258.