雲芝菌絲體發酵液凍乾粉之安全性探討

Investigations into the Acute Toxic and Mutagenic Activity of Freeze-dried Trametes Versicolor Mycelial Fermentation Broth Powder

目的：以三項基因毒及小鼠急毒性評估模式來初步探討雲芝菌絲體發酵液凍乾粉之安全性。方法：沙門氏菌回復突變試驗以五株試驗菌株於鼠肝萃物（S9）存在與否下各進行5個劑量組、1個陽性及1個陰性對照組共7組三重複試驗。體外哺乳類細胞染色體結構異常試驗是以CHO-K1倉鼠卵巢細胞，於S9存在與否下進行3個劑量試驗。囓齒類週邊血液微核試驗是以雄性ICR品系小鼠為試驗對象，共設置正、負對照組及低、中、高劑量組五組，每組各使用5隻動物進行試驗。在單次投予試驗物質後48小時及72小時後，分別評估小鼠週邊血液之網狀紅血球數及網狀紅血球中微核發生率。急毒LD50分析單一劑量（12g/kg b.w.）試驗物質經由口服投予ICR小鼠，各組10隻，雌雄鼠各半，連續觀察7日，動物即禁食犧牲解剖切片評估腎、肝、脾組織病理。結果：沙門氏菌回復突變試驗結果顯示，無論是否經過大鼠肝臟S9酵素代謝作用，皆不會引發沙門氏菌回復突變。體外哺乳類細胞染色體結構異常試驗結果亦呈現陰性反應。週邊血液之網狀紅血球和網狀紅血球中微核數目，相較於陰性對照組均無顯著性差異，顯示對囓齒類動物之週邊血液微核試驗為陰性反應。另在餵食12g/kg b.w.之雌、雄各五隻小鼠急毒性試驗，結果皆未死亡，顯示其LD50大於12g/kg b.w.。結論：雲芝發酵液凍乾粉未具基因毒性及口服急毒性（LD50>12g/kg b.w.）。

Purposes: The present study was conducted to assess the genotoxicity and acute toxicity of freezedried Trametes versicolor mycelial fermentation broth powder. Methods: Five different concentrations of Trametes versicolor mycelialf ermentation broth powder, including positive and negative controls, were tested in triplicate for mutagenicity using five Salmonella typhimurium strains both in the presence and absence of metabolic activation. In addition, in vitro chromosome aberration tests in Chinese Hamster Ovary (CHO-K1) cells in the presence and absence of a metabolic activation system were as performed. In the in vivo erythrocyte micronucleus assay, three doses of the test material were administered to male mice by oral gavage and the frequencies of reticulocytes/red blood cells and micronucleated reticulocytes/reticulocytes in the bone marrow were determined at intervals of 48 and 72 h following dosage. The acute toxicity was evaluated at 7 d after a single dose oral administration of Trametes versicolor mycelial fermentation broth powder (12 g/kg b.w.) to ten male and female ICR mice. All animals were sacrificed and followed by a full, detailed gross pathological examination of their kidneys, livers, and spleens. Results: Our results have indicated that Trametes versicolor mycelial fermentation broth powder did not significantly increase the number of revertant colonies in the bacterial reverse mutation test nor induce a higher frequency of aberrations in the chromosome aberration test. Moreover, no statistically significant Trametes versicolor mycelial fermentation broth powder-related increase was observed in the incidence of reticulocytes/red blood cells and micronucleated reticulocytes/reticulocytes. No animals died after a single oral administration of Trametes versicolor mycelial fermentation broth powder at 12g/kg b.w. Therefore, the LD50 of Trametes versicolor mycelial fermentation broth powder is greater than 12g/kg b.w./ day. Conclusions: Trametes versicolor mycelial fermentation broth powder is non-genotoxic in a three standard battery of tests and its oral LD50 is greater than 12g/kg b.w.