生命科学竞赛论文生命科学竞赛论文.doc

贺兰山紫蘑菇抗肿瘤活性研究 摘要：本项目以贺兰山紫蘑菇为材料，深度研究并分析其抗肿瘤、抗氧化活性及分子机理。通过95％乙醇渗漉提取，提取液浓缩后以石油醚、乙酸乙酯和正丁醇萃取获得萃取物，MTT法体外抗肿瘤活性检测初步结果显示石油醚、乙酸乙酯相抗肿瘤效果显著，经过大批量提取，硅胶柱层析、制备薄层层析分离，核磁谱图鉴定得到了3个化合物，抗肿瘤、抗氧化活性结果显示乙酸乙酯提取物抗氧化活性显著，从中分离所得的Moridolide有抗肿瘤活性，经过分子实验的条带分析，Moridolide对乳腺癌细胞的抗癌作用可能是因为作用于cyclin D1及VEGF-C从而影响细胞的增殖,在IL-8上有一定的作用说明其在一定程度内减弱了肿瘤细胞的恶性程度。 关键词：贺兰山紫蘑菇；抗肿瘤；提取；抗氧化；RT-PCR The anticancer constituents of Cortinarius rufo-olivaceus Abstract: to investigate of the anti-tumor active components, antioxidant activity and molecular biological characteristics of Cortinarius rufo-olivaceus, to provide a theoretical basis for the follow-up work. The dried C rufo-olivaceus were powdered and extracted with ethanol. The solvent was fully rotary evaporated under reduced pressure to yield a brown solid, suspended in water, extracted with petroleum ether (PE), EtOAc and n-BuOH successively, preliminary results of MTT assay in vitro show that PE extract and EtOAc extract have potent antitumor effects, 3 compounds were separated with Silica gel column chromatography and preparative chromatography, indentified with NMR spectrum. The antioxidant activity and antitumor activity results showed that the antioxidant activity of EtOAc extract is potent, the obtained compound Moridolide has antitumor activity, effects on cyclin D1 and VEGF-C to influence cell proliferation may account for the anticancer activity of Moridolide on breast cancer cells, it is certified that Moridolide also has certain effect on IL-8 to make sure that Moridolide can weakened the malignant degree of tumor cells through the analysis of experimental molecular bands. Key words: Cortinarius rufo-olivaceus ;antitumor; extract; antioxidant; RT-PCR 贺兰山紫蘑菇(Cortinarius rufo-olivaceus)为外生菌根菌，与青海云杉树的根系有着密切的共生关系[1]。它是生长在贺兰山自然保护区森林中的一种肉质滑厚、口感纯正、气味香醇、含有丰富的粗纤维、总糖、蛋白质、多种矿质元素以及17种游离氨基酸[2]的野生食用菌，被誉为“贺兰山珍”。初步研究显示，贺兰山紫蘑具有明显的抗肿瘤活性，但是其活性成分及抗性机制不明确，给该菌的深度开发带来障碍，课题计划对此进行深度研究，以期开发高附加值的产品。1 材料与方法 流程一:抗癌活性检测 95%乙醇渗漉提取 风干后水煎煮 流程二：抗瘤活性定位