黄河三角洲来源鹅绒藤内生真菌Aspergillus iizukae RD16次级代谢产物研究

Secondary metabolites of the endophytic fungus Aspergillus iizukae RD16 isolated from Cynanchum chinene in the Yellow River Delta

摘要

目的 探讨黄河三角洲来源鹅绒藤内生真菌Aspergillus iizukae RD16的次级代谢产物及细胞毒性。 方法 利用硅胶柱色谱、凝胶柱色谱和半制备高效液相等方法和技术对Aspergillus iizukae RD16大米发酵产物进行分离、纯化。使用NMR及质谱数据分析鉴定化合物的结构;采用CCK-8法评价化合物的细胞毒活性。 结果 从采自黄河三角洲的植物鹅绒藤中分离获得一株内生真菌RD16,鉴定为Aspergillus iizukae RD16;从其大米发酵产物中分离鉴定了7个单体化合物;分别为cytochalasin Z17(1)、cytochalasin Z16(2)、flavichalasine I(3)、aspochalasin J(4)、isorhodoptiometrin(5)、questin(6)、sulochrin(7)。其中化合物(1~4)为细胞松弛素类化合物,化合物(5和6)为蒽醌类化合物,化合物(7)为二苯甲酮类化合物。采用CCK-8法测试所有化合物对五株肿瘤细胞的毒性,其中化合物1对非小细胞肺癌A549细胞有显著的抑制活性,IC₅₀值为2.6 μmol/L;化合物2对A549细胞以及人结直肠腺癌RKO细胞均具有较强的抑制活性,IC₅₀值分别为3.0 μmol/L和2.8 μmol/L;化合物4对RKO细胞的IC₅₀值为5.6 μmol/L,对A549细胞有中等强度的细胞毒活性,其IC₅₀值为11.3 μmol/L。化合物5对SW480细胞有较强的抑制活性,IC₅₀值为5.8 μmol/L,对A549细胞有中等细胞毒活性,IC₅₀值为13.4 μmol/L。化合物6对A375以及A549细胞的IC₅₀值分别为8.5 μmol/L和3.9 μmol/L。 结论 黄河三角洲盐碱地来源植物内生真菌A.iizukae RD16可以产生结构多样的次级代谢产物。细胞松弛素类和蒽醌类化合物,在抗肿瘤先导化合物发掘方面具有一定应用潜力。

Abstract

Objective To investigate the secondary metabolites and cytotoxic properties of the endophytic fungus Aspergillus iizukae RD16, which was isolated from the plant Cynanchum chinense in the Yellow River Delta. Methods The fermentation products of the Aspergillus iizukae RD16 strain were isolated and purified using silica gel column chromatography, gel filtration chromatography and semi-preparative high performance liquid chromatography. The structures of the compounds were identified using NMR and mass spectrometry data analysis. The CCK-8 assay was used to evaluate the cytotoxic activity of the compounds. Results An endophytic fungus from Cynanchum chinene collected in the Yellow River Delta was identified as A.iizukae RD16. Seven monomeric compounds were isolated and identified in its rice fermentation products: cytochalasin Z17(1), cytochalasin Z16(2), flavichalasine I(3), aspochalasin J(4), isorhodoptiometrin(5), questin(6), and sulochrin(7). Compounds 1-4 belong to the cytochalasins, compounds 5 and 6 are anthraquinones, and compounds 7 is a benzophenone. The cytotoxic effects of all the compounds were tested against five human tumor cell lines using the CCK-8 assay. The results showed that compound 1 exhibited significant inhibitory activity against A549 non-small cell lung cancer cells, with an IC₅₀ value of 2.6 μmol/L. Compound 2 demonstrated potent inhibitory activity against both A549 cells and RKO human colorectal adenocarcinoma cells, with IC₅₀ values of 3.0 μmol/L and 2.8 μmol/L, respectively. Compound 4 exhibited moderate activity against A549 cells, with an IC₅₀ value of 11.3 μmol/L, and an IC₅₀ value of 5.6 μmol/L against RKO cells. Compound 5 displayed strong activity against SW480 cells(IC₅₀ value of 5.8 μmol/L)and moderate activity against A549 cells(IC₅₀ value of 13.4 μmol/L). Compound 6 exhibited IC₅₀ values of 8.5 μmol/L and 3.9 μmol/L against A375 cells and A549 cells, respectively. Conclusion The endophytic fungus Aspergillus iizukae RD16 which is found in the Yellow River Delta produces a variety of secondary metabolites. Among them, Cytochalasins and anthraquinones show potential for the discovery of lead compounds due to their cytotoxic activities.

关键词

黄河三角洲 / 次级代谢产物 / 内生真菌 / Aspergillus iizukae RD16 / 细胞毒活性

Key words

Yellow River Delta / Secondary metabolites / Endophytic fungi / Aspergillus iizukae RD16 / Cytotoxic activity

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基金资助

国家自然科学基金(81903537)

山东省医药卫生发展计划(202313050516)

山东省中医药科技发展计划(Z20244106)

滨州市“揭榜挂帅”技术攻关计划(2025JBGS007)