中国弯颈霉提取物抗氧化和抗肿瘤体外活性研究

Study on Anti-oxidant and Anti-tumor Activities in vitro of Tolypocladium sinense Extracts

中国弯颈霉(Tolypocladium sinense)分离自冬虫夏草,其生长最适温度25～28℃,最适pH6～7。在液体培养基(葡萄糖4%、蛋白胨0.5%、酵母膏1%、KH2PO40.1%、MgSO40.05%)中,经过28℃下摇床(150r/min)培养4～10d后,菌丝体生物量达8.49～9.73g/L。其菌丝体石油醚、乙酸乙酯、无水乙醇和蒸馏水提取率分别为3.51%、1.62%、4.84%、23.33%,菌丝体多糖提取率为5.31%。各提取物对DPPH自由基均具有清除作用,2mg/ml多糖具有较强的清除作用,清除率为49.13%;菌丝体多糖(0.08mg/ml)能显著地清除羟自由基,清除率为82.5%。MTT法检测发现乙酸乙酯提取物具有较强的抑制白血病细胞增殖作用;作用于K562和SHI-1细胞2d后,乙酸乙酯提取物IC50分别为26.55、17.75μg/ml;该提取物对正常细胞(人PBMC和小鼠骨髓细胞)的抑制作用较弱;AO/EB双染和TUNEL法检测提示菌丝体的乙酸乙酯提取物可以诱导K562细胞凋亡。上述实验结果表明,中国弯颈霉具有抗氧化和抗肿瘤方面的应用开发前景。

Tolypocladium sinense was isolated from the fruiting body of a wild Cordyceps sinensis. After growing for 4 to 10 days at 25 to 28 ℃ by shake culture (150 r/min) in a liquid medium (pH 6 to 7) containing glucose 4%, yeast extract 1%, peptone 0.5%, KH2PO4 0.1% and MgSO4 0.05%, the mycelial biomass of Tolypocladium sinense reaches 8.49 to 9.73 g/L. The mycelia extraction rates with petroleum, ethyl acetate, ethanol and hot water are 3.51%, 1.62%, 4.84% and 23.33%, respectively, and the extraction rate of myce...