首页|蛹虫草不同水提物体外抗白血病活性研究

蛹虫草不同水提物体外抗白血病活性研究

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蛹虫草(Cordyceps militaris是一种著名的药用菌,在东亚地区已被广泛用作中药材和滋补品,对多种肿瘤细胞有治疗作用.本研究探讨了蛹虫草液体培养的菌丝体(CMME)、发酵液(CMBE)和全发酵产物(菌丝体和发酵液,CMPE)的水提物对人(Homosapiens)白血病细胞Jurkat和K562的作用.四甲基偶氮唑蓝(methylthiazoletetrazolium,MTT)分析结果表明,这3种水提物对两种白血病细胞均有显著的抑制作用,在处理24h时已经比较明显,且呈剂量和时间依赖性.与CMBE相比,CMME和CMPE对人白血病细胞的抑制效果更为显著,但两者的差异则不显著.这3种水提物对两种细胞的半数抑制浓度(IC50值)分别为CMME<CMPE<CMBE.Annexin V-FITC/PI双标记流式细胞术检测结果表明,CMME和CMPE的抑制效应与细胞凋亡有关,且早期和晚期的Jurkat和K562细胞凋亡率与水提物浓度呈正比.同时,CMME和CMPE对Jurkat细胞的凋亡诱导效果相似,但前者能引起更多的K562细胞凋亡.整体上,3种水提物中以CMME对细胞的抑制效应和细胞凋亡作用较为明显.另外,通过Westem blot法检测抗凋亡因子Bcl-xL(B-cell lymphoma-extra large)的表达水平发现,处理48 h后,4.00 mg/mL的CMME能显著抑制Jurkat细胞中Bcl-xL蛋白的表达,而在高浓度CMME(8.00 mg/mL)的长时间处理(60 h)下,K562细胞中Bcl-xL蛋白的表达量略有下降.以上数据表明,蛹虫草水提物的抗肿瘤活性与细胞凋亡有关,且CMME在白血病治疗中比CMBE和CMPE更具有应用潜力.本研究首次比较了蛹虫草全发酵产物、发酵液和发酵菌丝体水提物对肿瘤细胞生长的抑制作用,蛹虫草3种水提物抗白血病活性的研究结果表明,其活性物质的产生主要为菌丝体内,而非分泌到细胞外.本研究结果为蛹虫草用于白血病治疗及其他相关研究提供基础数据,并为下一步探索其机理提供一定的参考资料.
Antileukemia Properties of Different Aqueous Extracts from Cordyceps militaris In vitro
As a well-known medicinal fungus,Cordyceps militaris has been widely used as a crude drug and folk tonic food in East Asia.It has been proved that C.militaris has the therapeutic actions against various kinds of cancer cells,while studies concerning the leukemia cells are not common,especially with fermentation products of this fungus.In this study,the effects of three aqueous extracts of C.militaris on human leukemia Jurkat and K562 cells were investigated.All aqueous extracts from C.militaris mycelium (CMME),fermentation broth (CMBE) and whole fermentation product (CMPE,extracts from both mycelium and fermentation broth) significantly inhibited the growth of Jurkat and K562 cells in a concentration-and time-dependent manner and the cytotoxic effects were clearly evident even from the early time point (24 h) using methylthiazoletetrazolium (MTT) assay.In comparison with CMBE,both CMME and CMPE showed a remarkably higher inhibitory effect on human leukemia cells,while no significant difference was observed between the two.Additionally,for the three extracts from liquid culture of C.militaris,IC50 value (inhibitory concentration for 50% growth inhibition) followed the ascending sequence:CMME<CMPE<CMBE.Further,the Annexin V-FITC/PI staining and flow cytometric analysis showed that the cytotoxic effects observed in response to the CMME and CMPE were associated with the induction of apoptotic cell death,and both early and late apoptosis rates were increased in a dose dependent manner in both Jurkat and K562 cells after the treatments.Both CMME and CMPE showed similar effects on the Jurkat cell apoptosis induction,although the former induced more K562 cells to undergo apoptosis than the latter.Generally,among the three extracts,CMME showed a stronger effect on cell viability and apoptosis than both CMBE and CMPE.In response to CMME,the level of anti-apoptosis B-cell lymphoma-extra large(Bcl-xL) expression was significantly inhibited in Jurkat cells after a relatively long time (48 h) at the concentration of 4.00 mg/mL,but slightly decreased in K562 cells (up to 60 h of treatment) at a higher concentration of 8.00 mg/mL.Taken together,the results here suggested that the anti-proliferative properties of C.militaris aqueous extracts were associated with the apoptotic induction and that CMME might have a great therapeutic potential in human leukemia treatment than the other two extracts.Antileukemia activities of the three aqueous extracts from C.militaris reported here indicated that the effective compounds were produced intracellularly rather than extracellularly.Further investigation may focus more on the mycelium production rather than fermentation broth.The results provide references for future study on application of C.militaris products in leukemia treatments and other related research.

Cordyceps militarisAqueous extractsHuman leukemia cellsAnticancer propertyApoptosis

席瑞娇、王文婧、陈吉龙、姚一建

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中国科学院微生物研究所,真菌学国家重点实验室,北京100101

中国科学院大学,北京100049

中国科学院微生物研究所,中国科学院病原微生物与免疫学重点实验室,北京100101

蛹虫草 水提物 白血病细胞 抗肿瘤活性 细胞凋亡

国家自然科学基金委面上项目国家重点基础研究发展计划(973计划)前期专项课题国家科技支撑计划课题科技基础性工作专项项目

311700172012CB1263082013BAD16B012012FY111600

2014

农业生物技术学报
中国农业大学 中国农业生物技术学会

农业生物技术学报

CSTPCDCSCD北大核心
影响因子:0.801
ISSN:1674-7968
年,卷(期):2014.22(7)
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