Antrodia cinnamomea Chang & Chou is a new, rare and expensive fungus first identified in 1994 in Taiwan. Our grant proposal plans to examine the safety and anti-genotoxic effects of Antrodia cinnamomea. We will determine the potential genotoxicity of Antrodia cinnamomea in female mice and their offspring using an established hprt gene mutation assay and micronuclei assay. After gavage ingestion of aqueous extracts of Antrodia cinnamomea from day 6 to day 18 of pregnancy, micornuclei in mouse polychromatic erythrocytes will be scored under fluorescent microscopy. The dams and pups will be sacrificed one month after delivery. The splenic lymphocytes will be isolated and cultured in the presence of the selective agents 6-thioguanine. The mutant colonies will be counted to determine hprt mutation frequency. These data will shed light on the genotoxicity potential of Antrodia cinnamomea and help us evaluate whether Antrodia cinnamomea is safe health food. Furthermore, the anti-genotoxic effects of Antrodia cinnamomea in mice will be examined by using the radiation-induced DNA damage and mutation animal model. The oxidative DNA damage, DNA strand break, micronuclei and hprt mutation frequency will be used as biomarkers to assess the protective effects of Antrodia cinnamomea administrated before or after irradiation. The oxidative DNA damage (8-hydroxy-2？H？H-deoxyguanosine) in the tissues will be analyzed using HPLC/ECD. The amounts of DNA strand breaks will be quantitated with alkaline COMET assay. Micornuclei in mouse polychromatic erythrocytes will be scored under fluorescent microscopy. The mononuclear cells from the mouse spleens will be isolated and the hprt mutation frequency in the T-lymphocytes will be examined in the presence of the selective agent 6-thioguanine. To further understand the underlying mechanism of the radio-protective effects of Antrodia cinnamomea, the level of oxygen free radicals in peripheral blood and the amounts of lipid peroxidation in mouse tissues will be analyzed by chemiluminescence detector and TBAR method, respectively. The data from this study will help us to elucidate whether the radioprotective effects of Antrodia cinnamomea is related to its protective effects against radiation-induced DNA damage, mutations, and lipid peroxidation.
