大蟬花菌種分離、鑑定及其液態培養菌絲體之基因毒性評估

Isolation, Identification and Genotoxicity Test of Cordyceps cicadae mycelium

徐瑞霞(Jui-Hsia Hsu) ； 葉淑幸(Shu-Hsing Yeh) ； 黃維茜(Wei-Chien Huang) ； 陳勁初(Chin-Chu Chen)

《檢驗及品保雜誌》 4卷3期 (2015/08) Pp. 114-127

https://doi.org/10.6573/JTQA.201508_4(3).03

摘要

大蟬花(Cordyceps cicadae)是傳統中藥材，具有多樣生理活性，其成分及功效與冬蟲夏草相似，菌絲體可工業化大量生產。本研究以自台灣山區所採集、分離之大蟬花為樣品，經鑑定確認後公開寄存，並以深層發酵培養菌絲體，再以三項基因毒性試驗來探討大蟬花菌絲體發酵液凍乾粉之安全性。結果顯示，在沙門氏桿菌回復突變測試中，不論在有無S9 代謝活化的條件下，在最高劑量5 mg/plate 以下各劑量組，均與陰性對照組不具顯著差異，顯示其對沙門氏菌不具致突變性。體外哺乳類細胞染色體結構異常試驗結果顯示，在三小時處理（含或不含S9 代謝活化混合物）以及十八小時處理（不含S9 代謝活化混合物）組別，所觀察到具染色體變異之細胞數，相較於陰性對照組皆無顯著差異，顯示在5 mg/mL 及其以下各濃度，對哺乳類細胞CHO-K1 不具有致染色體變異之基因毒性。在生體內哺乳類動物細胞微核測試結果亦顯示在5,000 mg/kg b.w. 及其以下各濃度組，皆不具誘發小鼠周邊血球細胞產生微核之能力；此外，所有測試劑量組的多染性紅血球比例和陰性對照組相較下皆無明顯差異，顯示試驗物質不會抑制小鼠之造血功能。綜合上述指出大蟬花菌絲體發酵液凍乾粉不具有基因毒性。

關鍵字

蟬花 ； 鑑定 ； 沙門氏桿菌回復突變試驗 ； 體外哺乳類細胞染色體結構異常試驗 ； 囓齒類週邊血液微核試驗

並列摘要

Cordyceps cicadae is an edible and traditional medicinal mushroom that possess multiple bioactivities. It has long been used as a desirable alternative for natural Ophiocordyceps sinensis and has therefore been mass reproduced for the increased market demand. In this research, we collected C. cicadae samples from the mountains of New Taipei City in Taiwan. These specimens were then sequenced, deposited at Bioresource Collection and Research Center at Food Industry Research and Development Institute and analyzed for genotoxicity using three standard battery of tests (Ames, chromosomal aberration, and bone marrow cell micronucleus tests). Results showed no cytotoxic or mutagenic potential in the Ames test at levels of up to 5 mg/plate. In the chromosomal aberration test, C. cicadae mycelium also showed no genetic toxicology when exposed to the CHO-K1 cells in the presence and absence of a metabolic activation system derived from rat liver S9 mix at levels of up to 5 mg/ml. Moreover, no statistically significant C. cicadae mycelium-related increase was observed in the bone marrow cell micronucleus test at levels of up to 5,000 mg/kg b.w.. Additionally, C. cicadae mycelium did not interfere with mouse bone marrow hematopoiesis evidenced by a no significant difference in the proportion of polychromatic erythrocytes in the peripheral blood. These results conclude that C. cicadae mycelium does not provoke mutagenicity and genotoxicity in these systems.

並列關鍵字

Cordyceps cicadae ； strain identification ； Ames test ； chromosomal aberration test ； bone marrow cell micronucleus test