华泽兰内生真菌Aspergillus puniceus次级代谢产物及其抗三阴性乳腺癌活性和作用机制  

Secondary metabolites of endophytic fungus Aspergillus puniceus from Eupatorium chinense and its anti-triple negative breast cancer activity and mechanism

作　　者：魏敏 陈莲 陈剑锋 金桂兰[2] 徐帮[2] 王倩 邹坤 程凡 刘呈雄 WEI Min;CHEN Lian;CHEN Jianfeng;JIN Guilan;XU Bang;WANG Qian;ZOU Kun;CHENG Fan;LIU Chengxiong(Hubei Key Laboratory of Natural Products Research and Development,College of Biological and Pharmaceutical Sciences,China Three Gorges University,Yichang 443002,Hubei,China;Yichang First People’s Hospital(People’s Hospital Affiliated to China Three Gorges University),Yichang 443002,Hubei,China;Yichang Humanwell Pharmaceutical Co.,Ltd,Yichang 443002,Hubei,China)

机构地区：[1]三峡大学生物与制药学院,天然产物研究与利用湖北省重点实验室,湖北宜昌443002 [2]宜昌市第一人民医院,三峡大学附属人民医院,湖北宜昌443002 [3]宜昌人福药业有限责任公司,湖北宜昌443002

出　　处：《华中师范大学学报（自然科学版）》2024年第2期206-213,共8页Journal of Central China Normal University：Natural Sciences

基　　金：湖北省中医药管理局中医药科研项目(ZY2023F083);天然产物研究与利用湖北省重点实验室(三峡大学)开放基金项目(2022NPRD03)。

摘　　要：从华泽兰根部分离得到一株内生真菌Aspergillus puniceus.采用正相硅胶柱色谱、Sephadex LH-20凝胶柱色谱及半制备高效液相色谱等方法,从该菌株的发酵液中分离得到8个结构类似的口山酮类化合物,通过现代波谱技术分别鉴定为3′,4′-hydrogenated austocystins A(1)、austocystin M(2)、austocystin F(3)、austocystin D(4)、austocystin B(5)、austocystin K(6)、austocystin A(7)、austocystin H(8).MTT结果显示化合物5对三阴性乳腺癌MDA-MB-231细胞具有较好的选择性细胞毒活性(IC_(50)为1.28μmol·L^(-1)).另外,流式细胞术检测化合物5作用于MDA-MB-231细胞后凋亡率的变化,相比于对照组,最高总凋亡率为69.7%.同时,AO/EB双荧光染色也证明化合物5能够诱导MDA-MB-231的细胞凋亡.JC-1线粒体膜电位检测进一步显示化合物5能够通过线粒体途径诱导三阴性乳腺癌MDA-MB-231的细胞凋亡.An endophytic fungus Aspergillus puniceus was isolated from the root fermentation of Eupatorium chinense.Eight structurally similar xanthones were isolated from the fermentation broth of the strain by normal phase silica gel column chromatography,Sephadex LH-20 gel column chromatography and semi-preparative high performance liquid chromatography.3′,4′-hydrogenated austocystins A(1),austocystin M(2),austocystin F(3),austocystin D(4),austocystin B(5),austocystin K(6),austocystin A(7),austocystin H(8)were identified by modern spectroscopic techniques.MTT assay showed that compound 5 had a preferably selective cytotoxicity against triple-negative breast cancer MDA-MB-231 cells with IC_(50) of 1.28μmol·L^(-1).In addition,flow cytometry was used to detect the change of apoptosis rate of MDA-MB-231 cells after treated with compound 5.Compared with the control group,the highest total apoptosis rate was 69.7%.Meanwhile,AO/EB double fluorescence staining also confirmed that compound 5 could induce apoptosis of MDA-MB-231 cells.Mitochondrial membrane potential detection of JC-1 further showed that compound 5 induced apoptosis in MDA-MB-231 cells through mitochondrial pathway.

关 键 词：华泽兰 Aspergillus puniceus 次级代谢产物 口山酮 三阴性乳腺癌 凋亡 

分 类 号：R917[医药卫生—药物分析学] R931.6[医药卫生—药学]