【作者】 刘崇玉；

【导师】 钱秀萍；

【作者基本信息】 上海交通大学 ， 制药工程， 2012， 硕士

【摘要】 植物内生真菌丰富多样的次级代谢产物是人们寻找新型抗菌、抗肿瘤天然药物的重要资源。本论文通过对植物内生真菌代谢物抗乳腺癌细胞(MDA)、肺癌细胞(A549)和胰腺癌细胞(PANC1)的活性筛选,以抗胰腺癌细胞活性较高的菌株为研究对象,进行了菌种鉴定和次级代谢产物的分离纯化、结构鉴定和活性检测,为深入研究提供有益的参考。本论文采用MTT法筛选了192株植物内生真菌的抗肿瘤细胞活性,88株植物内生真菌代谢物的肿瘤细胞生长抑制率在50%以上,其中2株内生真菌HCCB05974和HCCB06030具有较高的抗胰腺癌细胞活性。通过对HCCB05974和HCCB06030菌株的菌落形态、培养特征和ITS序列分析,分别归类为刺盘孢属和链格孢属。采用石油醚萃取、减压硅胶柱层析、HPLC制备等方法对菌株HCCB05974和HCCB06030的次级代谢产物进行分离纯化,并利用质谱和1H-NMR等相关波谱对化合物进行结构解析。HCCB05974来源的5个化合物鉴定为Nectriapyrone、3-吲哚甲酸乙酯、苯乙酸、对羟基苯乙酸和吲哚-3-乙酸乙酯,HCCB06030来源的6个化合物鉴定为Solaniol、dl-aloesol、Alternariol、Isoaltenuene、Altenuene和交链孢霉甲基醚。考察这些化合物对乳腺癌细胞(MDA)、肺癌细胞(A549)和胰腺癌细胞(PANC1)的活性,结果表明来源于刺盘孢属HCCB05974内生真菌的吲哚-3-乙酸乙酯具有弱抗肿瘤活性。更多还原

【Abstract】 The abundant and various metabolites from endophytic fungi have became an important resource of new kinds of natural drugs with anti-bacterial and anti-tumor activities. In this paper, the anti-tumor activities of metabolites from endophytic fungi were screened on the model of breast cancer cells (MDA), lung cancer cells (A549) and pancreatic cancer cells, and the strains which have higher anti-tumor activities were identified. The secondary metabolites were purified ,at the same time, the structure and anti-tumor activities of these compounds were studied.In this study, the metabolites of 192 kinds of endophytic fungal strains were studied by the methods of MTT, it was found that the inhibition growth rate of tumor cells were higher than 50% among the metabolites from 88 kinds of fungi and the metabolites from the strains of HCCB05974 and HCCB06030 had more activities against PANC1 cells. Based on the morphological, cultural characteristics and ITS sequences, the strains of HCCB0597 and HCCB06030 were respectively identified as Colletotrichum sp. and Alternaria sp. Secondary metabolites of these two strains were purified by the methods of petroleum ether extracting, silica gel column chromatography and HPLC preparation, according to the mass spectrometry and 1H-NMR spectral analysis ,the results showed that 5 kinds of compounds obtained from HCCB05974 were identified as nectriapyrone, 3-indole ethyl, phenylacetate, hydroxyphenyl acetic acid and indole-3-ethyl acetate ester, and 6 kinds of compounds from HCCB06030 were identified as solaniol, dl-aloesol, alternariol, isoaltenuene, altenuene and alternaria mold methyl ether. The effects of these compounds on the mold of MDA, A549 and PANC1 were studied and the indole-3-ethyl acetate had weak anti-tumor activity.更多还原