喜树内生真菌的分离及其抗肿瘤活性代谢产物的筛选方法

Isolation of endophytic fungi from Camptotheca acuminata and the screening method of antitumor secondary metabolite produced by the fungi

从喜树(CamptothecaacuminataDecne.)的根、枝条、叶和果实中分离纯化了48株内生真菌,通过对各个菌株的少量发酵培养,HPLC分析结合色谱峰紫外扫描检测方法,以紫外扫描图谱的相似性为依据,对喜树内生真菌产生喜树碱结构类似物进行初步筛选,并进一步以抑瘤实验确证其抗肿瘤活性。结果证明,以该方法筛选到的10个内生菌株中有7个菌株发酵液对HL 60细胞增殖具有显著的抑制活性。相对于常规的生物活性筛选,高效液相色谱结合色谱峰紫外光谱的方法,在药用植物内生真菌活性次生代谢产物筛选研究中具有快速、高效的特点。

The 48 strains of endophytic fungi were isolated from roots, branches, leaves and fruits of Camptotheca acuminata Decne. The secondary metabolites of these strains were screened for camptothecin and analog using the method of HPLC combined with the ultraviolet scanning spectrum according to their ultraviolet spectrums similar to camptothecins' and 10 strains which could produce camptothecin analog were acquired. Their antitumor activities were screened and 7 of them could evidently inhibit the growth of HL-60 cells in vitro. This results show that, compared with the ordinary biological screening method, the HPLC-UV screening method is fast, convenient and more suitable for small scale preliminary screening purpose.