巴西蘑菇原產於巴西，巴西蘑菇最常用來抑制癌症也被用來改善許多疾病，例如:糖尿病、高膽固醇、軟骨症。巴西蘑菇含有豐富的多醣體，這些多醣體被發表可增進免疫功能進而殺死癌細胞。雖然巴西蘑菇的好處很多，但它的生物特性及成分還沒有被完整的研究清楚。我們的研究就是去探討巴西蘑菇的抗氧化性，我們利用肺癌上皮細胞A549，骨癌上皮細胞Saos-2、老鼠纖維母細胞NIH3T3 和大鼠的心肌細胞H9c2 當作研究材料。這些細胞加入不同濃度的巴西蘑菇萃取液，再利用MTT 方法觀察細胞存活率。結果發現加入巴西蘑菇萃取液高達10 mg/ml 也不會影響細胞存活率，顯示說巴西蘑菇在10mg/ml 濃度下不具有細胞毒性。另外當細胞遭受到氧化性傷害時巴西蘑菇也顯示出保護作用。為了研究巴西蘑菇對細胞分子層面作用機制，我們利用NIH3T3 細胞加入巴西蘑菇萃取液並用過氧化氫(H2O2)來製造細胞的氧化性傷害，接著用流式細胞儀偵測反應性氧化物質(ROS)的數量。結果發現加入巴西蘑菇萃取液可減少ROS 的數量。再來我們利用彗星分析(comet assay)和南西方墨點法(south-western immunoblot)觀察到巴西蘑菇萃取液可減少H2O2 造成的氧化性DNA，即是8-oxoguanine 的量減少。因此我們的結果發現巴西蘑菇更進一步，本實驗利用硫酸銨鹽析沉澱法找到降低巴西蘑菇中降低ROS 的分餾物，但是我們目前尚未找到巴西蘑菇影響ROS 的分子機制，這將是未來主要的研究方向。

　　The Agaricus blazei Murill (ABM) mushroom is native to south-eastern Brazil and has been known for its medicinal properties. The mushroom is used to combat diabetes, hypercholesterolemia and osteoporosis. The protein-polysaccharide complex extracted from A. blazei Murill have previously been shown to have antitumor activity and stimulate immunity benefits. Nevertheless, scientific knowledge about its biological properties is still insufficient. In the present study, the antioxidant effects of A. blazei have been studied. By the analysis of MTT assay, the mouse fibroblast (NIH3T3), lung carcinoma epithelial cell (A549), osteosarcoma epithelial cell (Saos-2) and rat heart myocardium (H9c2) were treated with A. blazei. The results indicate the concentration (≦10 mg/ml) of A. blazei is no genotoxic. The aqueous extract dose (≦4 mg/ml) of A. blazei is able to protect from oxidative damage in all cells except, but not in Saos-2 cell. To investigate the molecular affect of A.blazei in NIH3T3 cell. We found the addition of A. blazei to cells reduces the amounts of reaction oxygen species (ROS), as demonstrated by flow cytometry. By comet assay and south-western immunoblot, we also found the DNA damage caused by hydrogen peroxide was decreased with treatment of A. blazei. The results suggest that extracts of Agaricus blazei Murill present antioxidant activity. We find out the fraction of A. blazei which can reduce ROS by salting out with ammonium sulfate. But we have not identified the mechanism which the level of ROS is reduced by A. blazei. It remains future investigation to elucidate the detail mechanism.

Ahn WS, Kim DJ, Chae GT, Lee JM, Bae SM, Sin JI, Kim YW, Namkoong SE, Lee IP.2004. Natural killer cell activity and quality of life were improved by consumption of a mushroom genic extract,Agaricus blazei Murill Kyowa, in gynecological cancer patients undergoing chemotherapy. Internationsl Journal of Gynecological Cancer. 14: 589–594.

Ames BN, Shigenaga MK and Gold LS. 1993. DNA lesion, induible DNA repair, and cell division: Three key factors in mutagenesis and carcinogenesis. Environmental Health Perspectives. 93: 35-44.

Barbisan LF, Miyamoto M, Scolastici C, Salvadori, DMF, Ribeiro LR, Eira AF, de Camargo, JLV. 2002. Influence of aqueous extract of Agaricus blazei on rat liver toxicity induced by different doses of diethylnitrosamin. Journal of Ethnopharmacology.83: 25–32.

Barbisan LF, Spinardi-Barbisan AL, Moreira EL, Salvadori DM, Ribeiro LR, da Eira AF, de Camargo, JL. 2003a. Agaricus blazei (Himematsutake) does not alter the development of rat diethylnitrosamine-initiated hepatic preneoplastic foci. Cancer Science. 94: 188–192.

Barbisan LF, Scolastici C, Miyamoto M, Salvadori DMF, Ribeiro LR, da Eira AF, de Camargo JLV. 2003b. Effects of crude extracts of Agaricus blazei on DNA damage and on rat liver carcinogenesis induced by diethylnitrosamine. Genetics and Molecular Research. 2: 295–308.

Bellini MF, Giacomini NL, Eira AF, Ribeiro LR, Mantovani MS. 2003. Anticlastogenic effect of aqueous extracts of Agaricus blazei on CHO-K1 cells, studying different developmental phases of the mushroom. Toxicology in Vitro. 17: 465–469.

Cohen SM and Ellwein LB. 1991. Genetic errors, cell proliferation, and carcinogenesis. Cancer Research. 51: 6493-6505.

Chang R. 1996. Functional properties of edible mushrooms. Nutrition Reviews. 54: 11–15.

Chen L, Shao HJ, Su YB. 2004. Coimmunization of Agaricus blazei Murill extract with hepatitis B virus core protein through DNA vaccine enhances cellular and humoral immune responses. International immunopharmacology. 4: 403–409.

Chris JN and Ian DH. 2001. Cellular responses to DNA damage. Annual Review of Pharmacology and Toxicology. 41: 367-401.

Coussens LM, Werb Z. 2002. Inflammation and cancer. Nature. 420:860-867.

Delmanto RD, Lima PLA, Sugui MM, Eira AF, Salvadori DMF, Speit G, Ribeiro LR. 2001. Antimutagenic efffect of Agaricus blazei Murrill mushroom on the genotoxicity induced by cydophosphamide. Mutation Research. 496: 15–21.

Dong Q, Yao J, Yang XT, Fang JN. 2002. Structural characterization of a water-soluble beta-D-glucan from fruiting bodies of Agaricus blazei Murill. Carbohydrate Research. 337: 1417–1421

Ebina T, Fujimiya Y. 1998. Antitumor effect of a peptide– glucan preparation extracted from Agaricus blazei in a double grafted tumor system in mice. Biotherapy. 11: 259–265.

Fujimiya Y, Suzuki Y, Oshiman K, Kobori H, Moriguchi K, Nakashima H, Matumoto Y, Takahara S, Ebina T, Katakura R. 1998. Selective tumoricidal effect of soluble proteoglucan extracted from the basidiomycete, Agaricus blazei Murill, mediated via natural killer cell activation and apoptosis. Cancer Immunology, Immunotherapy. 46:147-159.

Fujimiya Y, Suzuki Y, Katakura R, Ebina T. 1999. Tumor-specific cytocidal and immunopotentiating effects of relatively low molecular weight products derived from the basidiomycete Agaricus blazei Murill. Anticancer Research. 19: 113-118.

Guterrez ZR, Mantovani MS, Eira AF, Ribeiro LR, Jordao BQ. 2004. Variation of the antimutagenicity effects of water extracts of Agaricus blazei Murrill in vitro. Toxicology in Vitro. 18: 301–309.

Hussain SP, Hofeseth LJ, Harris CC. 2003. Radical causes of cancer. Nature Review Cancer. 3:276-285.

Itoh H, Ito H, Amano H, Noda H. 1994. Inhibitory action of a (1-6)-ß-D-glucan-protein complex (F III-2-b) isolated from Agaricus blazei Murill (“Himematsutake”) on Meth A fribrosarcoma- bearing mice and its antitumor mechanism. Japanese Journal of Pharmacology. 66: 265-271.

Ito H, Shimura K, Itoh H, Kawabe M. 1997. Antitumor effects of a new polysaccharide-protein complex (ATOM) prepared from Agaricus blazei (Iwade Strain 101) “Himematsutake” and its mechanism in tumor-bearing mice. Anticancer Research. 17: 277-284.

Izawa S, Inoue Y. 2004. A screening system for antioxidants using thioredoxin-deficient yeast: discovery of themostable antioxidant activity from Agaricus blazei Murill. Applied Microbiology Biotechnology. 64: 537-542.

Kaneno R, Fontanari LM, Santos SA, Di Stasi LC, Rodrigues Filho E, Eira AF. 2004. Effects of extracts from Brazilian sun-mushroom (Agaricus blazei) on the NK activity and lymphoproliferative responsiveness of Ehrlich tumor-bearing mice. Food and Chemical Toxicology. 42:909-916.

Kasai H, He LM, Kawamura M, Yang PT, Deng XW, Munkanta M, Yamashita A, Terunuma H, Hirama M, Horiuchi I, Natori T, Koga T, Amano Y, Yamaguchi N, Ito M. 2004. IL-12 production induced by Agaricus blazei fraction H(ABH) involves Toll-like receptor(TLR). Evidence Based Complementary and Alternative Medicine. 3:259-267.

Kawagishi H, Katsumi R, Sazama T, Mizuno T, Hagiwara T, Nakamura T. 1988. Cytotoxic steroids from the mushroom Agaricus blazei. Phytochemistry. 27: 51–58.

Kawagishi H, Inagaki R, Kanao T, Mizuno T, Shimura K, Ito H, Hagiwara T, Nakamura T. 1989. Fractionation and antitumor activity of the water-insoluble residue of Agaricus blazei fruiting bodies. Carbohydrate Research. 186: 267–273.

Kawagishi H, Kanao T, Inagaki R, Mizuno T, Shimura K, Ito H, Hagiwara T, Nakamura T. 1990. Formolysis of a potent antitumor (1–6) ß-D-glucan-protein complex from Agaricus blazei fruiting bodies and antitumor activity of the resulting products. Carbohydrate Polymers. 12: 393–403.

Kawagishi H, Nomura A, Yumen T, Mizuno T, Hagiwara T, Nakamura T. 1998. Isolation and properties of a lectin from the fruiting bodies of Agaricus blazei. Carbohyhrdrate Research. 183: 150-154.

Kimura Y, Kido T, Takaku T, Sumiyoshi M, Baba K. 2004. Isolation of an anti-angiogenic substance from Agaricus blazei Murill: its antitumor and antimetastatic actions. Cancer Science. 95: 758-764.

Kimura Y. 2005. New anticancer agents: in vitro and in vivo evaluation of the antitumor and antimetastatic actions of various compounds isolated from medicinal plants. In Vivo 9: 37-60.

Kuroiwa Y, Nishikawa A, Imazawa T, Kanki K, Kitamura Y, Umemura T, Hirose M. 2005. Lack of subchronic toxicity of an aqueous extract of Agaricus blazei Murill in F344 rats. Food and Chemical Toxicology. 43: 1047-1053.

Lee YL, Kim HJ, Lee MS, Kim JM, Han JS, Hong EK, Kwon MS, Lee MJ. 2003. Oral administration of Agaricus blazei (H1 strain) inhibited tumor growth in a sarcoma 180 inoculation model. Experimental Animals. 52: 371–375.

Luiz RC, Jordao BQ, Eira AF, Ribeiro LR, Mantovani MS. 2003. Mechanism of antimutagenicity of Agaricus blazei Murrill mushroom organic extracts in wild type CHO (K1) and repair deficient (xrs5) cells by chromosome aberration and sister chromatid exchange assays. Mutation Research. 528: 75–79.

Machado MP, Filho ER, Terezan AP, Ribeiro LR, Mantovani MS. 2005. Cytotoxicity, genotoxicity and antimutagenicity of hexane extracts of Agaricus blazei determined in vitro by the comet assay and CHO/HGPRT gene mutation assay. Toxicology in Vitro. 19: 533-539.

Martindale JL, Holbrook N J. 2002. Cellular response to oxidative stress: signaling for suic ide and survival. Journal of Cellular Physiology. 174: 27-34.

Martins de Oliveira J, Jordao BQ, Ribeiro LR, Ferreira da Eira A, Mantovani MS. 2002. Anti-genotoxic effect of aqueous extracts of sun mushroom (Agaricus blazei Murill lineage 99/26) in mammalian cells in vitro. Food and Chemical Toxicology 40: 1775–1780

Menoli RCRN, Mantovani MS, Ribeiro LR, Speit G, Jordao BQ. 2001. Antimutagenic effects of the mushroom Agaricus blazei Murrill extracts on V79 cells. Mutation Research. 496: 5–13.

Mizuno M, Morimoto M, Minato K, Tsuchida H. 1998. Polysaccharides from Agaricus blazei stimulate lymphocyte T-cell subsets in mice. Bioscience Biotechnology and Biochemistry. 62: 434–437.

Mizuno, T. 2002. Medicinal properties and clinical effects of culinarymedicinal mushroom Agaricus blazei Murrill stimulate (Agaricomycetideae). International Journal of Medicinal Mushrooms. 4: 312–399.

Nakajima A, Ishida T, Koga M, Takeuchi T, Mazda O, Takeuchi M. 2002. Effect of hot water extract from Agaricus blazei Murill on antibody-producing cells in mice. International Immunopharmacology. 2: 1205–1211

Ohno N, Furukawa M, Miura NN, Adachi Y, Motoi M, Yadomae T. 2001. Antitumor beta-glucan from the cultured fruit body of Agaricus blazei. Biologica l and Pharmaceutical Bulletin. 24: 820–828.

Oliveira JM, Jordao BQ, Ribeiro LR, Eira AF, Mantovani MS. 2002. Anti-genotoxic effect of aqueous extracts of sun mushroom (Agaricus blazei Murill lineage 99/26) in mammalian cells in vitro. Food and Chemical Toxicology 40: 15–20.

Ooi VEC, Liu F. 2000. Immunomodulation and anti-cancer activity of polysaccharide-protein complexes. Current Medicinal Chemistry. 7:715-729.
Oshiman K, Fujimiya Y, Ebina T, Suzuki I, Noji M. 2002. Orally administered beta-1,6-D-polyglucose extracted from Agaricus blazei results in tumor regression in tumor-bearing mice. Planta Medica. 68:610–614.

Pinheiro F, Faria RR, de Camargo JL, Spinardi-Barbisan AL, da Eira AF, Barbisan LF. 2003. Chemoprevention of preneoplastic liver foci development by dietary mushroom Agaricus blazei Murrill in the rat. Food and Chemical Toxicology. 41: 1543–1550.

Rota C, Fann YC, Mason RP. 1999. Phenoxyl free radical formation during the oxidation of the fluorescent dye 2’,7’-Dichlorofluorescein by horseradish peroxidase. The Journal of Biological Chemistry. 274:28161-28168.

Satoh MS and Lindahl T. 1994. Enzymatic repair of oxidative DNA damage. Cancer Research. 54: 1899-1901.

Sander CS, Chang H, Hamm F, Elsner P, Thiele JJ. 2004. Role of oxidative stress and the antioxidant network in cutaneous carcinogenesis. International Journal of Dermatology 43: 326-335.

Shimizu S, Kitada H, Yokota H, Yamakawa J, Murayama T, Sugiyama K, Izumi H, Yamaguchi N. 2002. Activation of the alternative complement pathway by Agaricus blazei Murill. Phytomedicine. 9:536–545.

Sorimachi K, Akimoto K, Ikehara Y, Inafuku K, Okubo A, Yamazaki S. 2001a. Secretion of TNF- alpha, IL-8 and nitric oxide by macrophages activated with Agaricus blazei Murrill fractions in vitro. Cell Struture and Function. 26: 103–108.

Sorimachi K, Ikehara Y, Maezato G, Okubo A, Yamazaki S, Akimoto K, Niwa A. 2001b. Inhibition by Agaricus blazei fractions of cytopathic effect induced by western equine encephalitis (Wee) virus on Vero cells in vitro. Bioscience Biotechnology and Biochesmitry. 65:1645–1647.

Takasaburo E, Yoshiaki F. 1998. Antitumor effect of peptide-glucan preparation extracted from Agaricus blazei in a double-grafted tumor system in mice. Biotherapy Dordrecht. 11: 259–265.

Takeda Y, Togashi H, Shinzawa H, Miyano S, Ishii R, Karasawa T, Takeda Y, Saito T, Saito K, Haga H, Matsuo T, Aoki M, Mitsuhashi H, Watanabe H, Takahashi T. 2000. Spontaneous regression of hepatocellular carcinoma and review of literature. Journal ofGastroenterology and Hepatology 15: 1079–1086.

Takaku T, Kimura Y, Okuda H. 2001. Isolation of an antitumor compound from Agaricus blazei Murill and its mechanism of action. The Journal of Nutrition. 131: 1409-1413.

Talorete TPN, Isoda H, Maekawa T. 2002. Agaricus blazei (class Basidiomycotina) aqueous extract enhances the expression of c- Jun protein in MCF7 cells. Journal of Agricultural and Food Chemistry 50: 5162–5166