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目的分离鉴定阿维链霉菌I06A-01716发酵液中抗植物病原真菌活性成分,并进行抗真菌、抗肿瘤和免疫抑制活性研究。方法在抗真菌活性指导下,采用硅胶柱层析、凝胶柱层析、HPLC等分离手段对活性组分进行分离;通过高分辨质谱,1D-NMR和2D-NMR对其结构进行鉴定;采用琼脂平板纸片法,进行抗植物病原真菌的最低抑制浓度测定;采用MTT法检测其对肿瘤细胞生长的抑制作用;采用流式细胞仪测定其对细胞周期的影响。结果分离得到2个活性组分:1716-1和1716-2。1716-1和1716-2对人肝癌细胞HepG2的IC50值分别为2.42μg/mL和3.42μg/mL;对人口腔上皮癌细胞KB的IC50值分别为1.64μg/mL和6.73μg/mL。细胞周期研究表明,随着1716-2的浓度增加,在KB细胞中,G2/M期比例由对照组的15.23%逐步升高到36.53%;在HepG2细胞中,G2/M期比例由对照组的4.39%升高到35.78%。结论1716-1和1716-2结构分别与寡霉素B和寡霉素A一致。1716-1和1716-2有抗多种植物病原真菌活性。1716-1和1716-2在较低浓度下能够抑制人肝癌细胞HepG2和人口腔上皮癌细胞KB生长。1716-2使HepG2细胞和KB细胞阻滞在G2/M期。
OBJECTIVE To isolate and identify the active constituents of the phytopathogenic fungi in the fermentation broth of Streptomyces avermitilis I06A-01716 and to study their antifungal, antitumor and immunosuppressive activities. Methods Under the guidance of antifungal activity, the active components were isolated by silica gel column chromatography, gel column chromatography and HPLC separation. Their structures were identified by high resolution mass spectrometry, 1D-NMR and 2D-NMR. Agar slab paper method to determine the minimum inhibitory concentrations against phytopathogenic fungi; MTT assay was used to detect the inhibitory effect on the growth of tumor cells; and the cell cycle was measured by flow cytometry. Results Two active components were isolated: IC50 values of HepG2 and HepG2 of 1716-1 and 1716-2.1716-1 and 1716-2 were 2.42μg / mL and 3.42μg / mL, respectively. Hepatocellular carcinoma cells The IC50 values for KB were 1.64 μg / mL and 6.73 μg / mL, respectively. Cell cycle studies showed that with the increase of the concentration of 1716-2, the percentage of G2 / M phase in KB cells gradually increased from 15.23% to 36.53% in control group. In HepG2 cells, the proportion of G2 / M phase was increased from control group 4.39% to 35.78%. Conclusion The structures of 1716-1 and 1716-2 are consistent with oligomycin B and oligomycin A, respectively. 1716-1 and 1716-2 have activity against a variety of plant pathogenic fungi. 1716-1 and 1716-2 inhibited the growth of human hepatoma HepG2 cells and human oral epithelial carcinoma KB cells at lower concentrations. 1716-2 blocked HepG2 cells and KB cells at the G2 / M phase.