1株具有抗肝癌活性的狭叶柴胡内生真菌的鉴定

Identification of an endophytic fungus of Bupleurum scorzonerifolium Willd. with anti-hepatoma activity

目的对1株具有抗肝癌活性的狭叶柴胡内生真菌CHS4进行菌种鉴定,并对其抗肝癌活性进行初步检测。方法采用传统分类学方法及电子显微镜观察对CHS4进行形态鉴定,并采用真菌鉴定通用引物V9D与LS266对其基因组DNA进行扩增得到ITS序列,将已经测得的序列结果与Gen Bank中已知序列进行Blast分析,从而进行分子鉴定。利用MTT法对菌CHS4的乙醇粗提物进行抗肝癌活性检测。结果内生真菌CHS4经鉴定为链格孢属真菌互隔交链孢霉(Alternaria alternata);该菌株发酵液的乙醇粗提物对肝癌细胞HepG-2的抑制作用与其浓度成正相关,且IC50值为75.11μg/m L。结论从狭叶柴胡中分离到一株具有抗肝癌作用的内生真菌CHS4,可作为生产抗肝癌药物的候选资源。

Objective To identify an endophytic fungus of Bupleurum scorzonerifolium Willd. with antihepatoma activity and study its anti-hepatoma activity as well. Methods By using the traditional taxonomic methods, the identification of morphological characters were performed. Primers V9D and LS266 were used to amplify the ITS sequence from genome DNA of CHS4. The amplified sequence was compared with known sequences in Blast of NCBI. Anti-hepatoma activity was detected by MTF assay. Results Fungus CHS4 from B. scorzonerifolium Willd., was classified as Alternaria alternate by the morphology and ITS identification. The ethanol crude extract of CHS4 had the ability to inhibit HepG-2 cell growth. There was a positive correlation between inhibition effect and concentration and IC50 value was 75.11 μg/mL. Conclusion The strain CHS4 with anti-hepatoma activity can be regarded as alternative resource for producing anti-hepatoma drugs.