以艾姆氏測驗法評估破壁靈芝基因毒性

Evaluation of Genotoxicity of Wall-broken Spores of Ganoderma lucidium in the Ames Test

陳筠臻(Yun-Jen Chen) ； 吳銘芳(Ming-Fang Wu) ； 施勇綸(Yung-Luen Shih) ； 李慶孝(Ching-Hsiao Lee) ； 賴珊湖(Shan-Hu Lai) ； 周建聖(Jason Chou) ； 周育如(Yu-Ru Chou) ； 常傳訓(Chuan-Hsun Chang) ； 呂旭峰(Hsu-Feng Lu)

《Journal of Biomedical & Laboratory Sciences》 25卷3期 (2013/09) Pp. 98-104

摘要

本研究是以長庚生物科技公司所生產的破壁靈芝孢子樣品，藉由沙門氏菌基因逆突變測試其突變性。測試濃度共五種，分別為3 mg/ml，6 mg/ml，12 mg/ml，25 mg/ml，50 mg/ml。在兩種不同條件下（有或無S9的催化），以plate incorporation方法對五種菌株（TA97、TA98、TA100、TA102、TA1535）進行逆突變測試，實驗皆三重複。結果計算每一個培養基所含菌落數。不含S9之非活化組，陽性組之致變物濃度為1μg/plate 4-nitroquinoline-N-oxide（TA97及TA98），濃度為8μg/plate mitomycin c（TA102），以及濃度為1μg/plate sodium azide（TA100及TA1535）。含S9之活化組，陽性組之致變物分別濃度為2μg/plate Benzo[a]pyrene（TA98及TA102）與濃度為8μg/plate 2-aminoanthracene（TA97、TA100及TA1535）。無論是否經過S9處理，五種細菌生長結果皆不受影響，且菌落數與濃度不呈劑量依賴性。破壁靈芝孢子在代謝活化前後處理下，對TA97、TA98、TA100、TA102及TA1535所引發之基因毒性並無顯著之增加。

關鍵字

破壁靈芝孢子 ； 沙門氏菌基因逆突變

並列摘要

This study was conducted in order to assess the safety and tolerability of wall-broken spores of Ganoderma lucidium obtained from Chang Gung Biotechnology Corporation, Ltd. in general mutagenicity studies by Ames tests in vitro. For the preparation of Ganoderma lucidium dose levels for Ames test, a series of concentrations was prepared from stock solution by dilution, namely 3 mg/ml, 6 mg/ml, 12 mg/ml, 25 mg/ml and 50 mg/ml. Bacterial strains used were Salmonella typhimurium TA97, TA98, TA100, TA102 and TA1535. The AMES test applied the plate incorporation method, without or with S9 metabolic activation. For a proper estimate of variation, triplicate plating was used at each dose level. After the incubation period, the number of reverting colonies per plate was counted. The positive control without S9 fraction consisted of 1 μg/plate of 4-nitroquinoline-N-oxide for TA97 and TA98 strains; 8 μg/plate of mitomycin C for TA102 strain; and 1 μg/plate of sodium azide for TA100 and TA1535 strain; for any with S9 fraction, 2 μg/plate of benzo[a]pyrene was used for TA98 and TA102 strains, and 8μg/plate 2-aminoanthracene for TA97, TA100 and 1535 strains. Compared to the negative control, the Ganoderma lucidium solutions with S9 or without S9 did not affect bacterial growth. There were no dose-dependent increases or decreases in the number of revertant colonies neither with nor without metabolic activation. Generally, mutagenicity was negative in all strains with and without the S9 mix.

並列關鍵字

wall-broken spores of Ganoderma lucidium ； Ames tests