北蟲草與蜜環菌複方液態膳食補充品之安全性評估

Abstract

子囊菌屬的Cordyceps包含超過五百種的真菌，其中北蟲草 (Cordyceps militaris) 在傳統中草藥及西方研究中受到重視。許多研究發現北蟲草中的蟲草素、腺苷及多醣類物質對人體具抗憂鬱、免疫調節、抗發炎、抗腫瘤、抗腫瘤轉移、抗疲勞、抗氧化、降血脂等功效，因此具有開發成保健食品之潛力。蜜環菌 (Armillaria mellea) 是一種食用菌，具有抗氧化、抗發炎和免疫調節之活性，且具有誘導人體樹突細胞的成熟之功效，長期以來被用作傳統的中草藥。而蜜環菌與天麻之間有共生關係，許多研究表明蜜環菌具有與天麻相似的藥理學性質，也說明天麻與共生真菌的蜜環菌具有相似的功效。再加上天麻的產量較少以及生長速度緩慢，因此可考慮以蜜環菌作為天麻的替代品，具有開發之潛力。本研究以北蟲草、蜜環菌以及其他具安神作用之中藥材（百合、佛手柑、烏梅與洛神花）製成之複方液態膳食補充品為受試物質，且由於本液態膳食補充品尚未通過健康食品認證，依照衛福部食藥署健康食品安全性評估方法規定，需進行第二類之基因毒性與28天亞急毒性試驗，以評估本液態膳食補充品是否會對生物體造成基因毒性及亞急毒性傷害，並為其提供日後食用安全性之參考依據。由Ames test實驗結果顯示，受試物質在0.3-5 mg/plate劑量下對Salmonella typhimurium TA98、TA100、TA102、TA1535及TA1537菌株之回復突變菌數，與負控制組相比均無顯著統計上差異。體外哺乳動物染色體異常分析試驗結果顯示，各試驗劑量組之總變異率與負控制組相比皆無統計上顯著差異。在微核試驗中的結果顯示，各試驗劑量下之網狀紅血球比例以及微核細胞發生率和負控制組相比均無統計上之顯著差異。另外，於28天亞急毒性試驗中，Sprague Dawley (SD) 大鼠經本液態膳食補充品管餵後並無任何明顯的臨床症狀或死亡情形，且在血液學檢查、血清生化值分析、組織病理切片及尿液檢驗結果均無明顯與受試物質相關之不良反應及病變情形。由上述結果判斷，北蟲草、蜜環菌、百合、佛手柑複方之液態膳食補充品不會造成基因毒性，亦不會在SD大鼠中產生亞急毒性，故本試驗之無毒性顯示劑量 (no-observed-adverse -effect level, NOAEL) 為大於高劑量組別 (25 ml/kg bw/day) 之濃度。

Cordyceps militaris (CM) has been increasingly viewed as a substitute for Ophiocordyceps sinensis because of not only the relative species but also their similar active components. Many studies have reported that cordycepin, glucosides and polysaccharides in CM had anti-depression, immune regulation, anti-inflammatory, anti-tumor, anti-tumor metastasis, anti-fatigue, antioxidation, blood triglycerides reduction effects. Thus, it has the potential to develop a human health food. Armillaria mellea (AM) is an edible fungus that has antioxidant, anti-inflammatory and immunomodulatory activities and can induce the maturation of human dendritic cells. AM has been traditionally used as Chinese herbal medicine. There is a symbiotic relationship between AM and Gastrodia elata Blume. Many studies have shown that AM had similar pharmacological properties to G. elata, and it also shows that G. elata has similar effects to AM. AM can be considered as a substitute for G. elata, which has the potential for development of related dietary supplement. In this study, the liquid dietary supplement is made from CM, AM and two medicinal herbs (Lilium lancifolium Thunb. leaf and Citrus medica var. sarcodactylus). To assess the safety for human consumption I performed genotoxicity and 28-day sub-acute toxicity studies of the supplement which belongs to category 2 according to the regulations of Health Food Safety Assessment announced by the Ministry of Health and Welfare in Taiwan. The result of Ames test showed that the treatment with supplement at 0.3-5.0 mg/plate did not significantly increase in revertant colonies of Salmonella typhimurium TA98, TA100, TA102, TA1535 and TA1537 strains. The result of in vitro mammalian chromosome aberration test showed that the treatment with supplement at 0.3-5.0 mg/ml did not increase in chromosome aberrations on Chinese hamster ovary cells (CHO-K1). The result in the micronucleus test indicated that there was no statistically significant difference in the proportion of reticulocyte and the incidence of micronuclei between the supplement treated groups and the negative control group. Moreover, in the 28-day subacute toxicity test, there were no obvious clinical symptoms and death occurred in the SD rats fed with the supplement, and showed no significant adverse reactions and lesions associated with the test substance in hematology, serum biochemistry, urinalysis, and histopathological examination. Based on above results, it could conclude that the supplement did not cause genotoxicity and subacute toxicity, Therefore, the no-observed-adverse-effect level (NOAEL) was higher than the high-dose group which is 25 ml/kg bw/day liquid dietary supplement of Cordyceps militaris and Armillaria mellea formula.