川芎内生真菌Fusarium proliferatum的化学成分  

Chemical constituents of the endophytic fungus Fusarium proliferatum from Ligusticum chuanxiong Hort.

作　　者：旷歧轩 谭璐 罗禹 冯丹[1] 周煜 郭大乐[1] 任波[1] 邓赟[1] KUANG Qixuan;TAN Lu;LUO Yu;FENG Dan;ZHOU Yu;GUO Dale;REN Bo;DENG Yun(Key Laboratory of Standardization of Chinese Herbal Medicine of Ministry of Education,State Key Laboratory Breeding of Systematic Research Development and Utilization of Chinese Medical Resources,Co-founded by Sichuan Province and Ministry of Science and Technology,Pharmacy Department,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;School of Pharmacy,Southwest Medical University,Luzhou 646000,China)

机构地区：[1]成都中医药大学药学院,中药材标准化教育部重点实验室,四川省中药资源系统研究与开发利用重点实验室-省部共建国家重点实验室培育基地,成都611137 [2]西南医科大学药学院,泸州646000

出　　处：《应用与环境生物学报》2021年第5期1318-1325,共8页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然科学基金面上项目(81973460);四川省科技厅应用基础研究项目(2018JY0186);四川省教育厅科研项目(18ZA0191);成都中医药大学杏林学者学科人才科研提升计划(BSH2018008)资助。

摘　　要：研究川芎内生真菌Fusarium proliferatum的次生代谢产物及其细胞毒活性.采用硅胶柱层析、中压反相硅胶柱色谱、制备液相色谱等方法从该菌发酵产物的乙酸乙酯萃取相中分离得到23个化合物.通过波谱技术分析鉴定化合物结构,分别为7-氧代-β-谷甾醇(1),7-氧代-β-豆甾醇(2),过氧化麦角甾醇(3),9,11-脱氢过氧化麦角甾醇(4),5α,6α-环氧-3β-麦角甾-22-烯-7-酮(5),5α,6α-环氧-3β,8β,14α-羟基-5α-麦角甾-22E-烯-7-酮(6),3β-羟基-(22E,24R)-麦角甾-5,8,22-烯-7-酮(7),(4S,17R)-4-羟基-17-甲基甾醇(8),(3β,5α,6β,22E)-麦角甾-7,22-二烯-3,5,6-三醇(9),白僵菌素(10),对羟基苯甲醛(11),环-(L-脯氨酸-L-甘氨酸)(12),环-(甘氨酸-D-苯丙氨酸)(13),环-(2-羟基-脯氨酸-甘氨酸)(14),环-(L-苯丙氨酸-丙氨酸)(15),9-十八碳烯酸-2’,3’-二羟基丙酯(16),环-(8α-羟基-D-脯氨酸-L-苯丙氨酸)(17),环-(8β-羟基-L-脯氨酸-D-苯丙氨酸)(18),环-(8α-羟基-L-脯氨酸-L-苯丙氨酸)(19),环-(L-脯氨酸-L-亮氨酸)(20),环-(L-脯氨酸-D-苯丙氨酸)(21),环-(L-脯氨酸-L-苯丙氨酸)(22),白僵菌酮(23).采用MTT法测试化合物1-7和化合物10的细胞毒活性,结果显示化合物3和化合物5表现出对MV4-11细胞株的抑制作用,IC50值分别为31.73±2.08和14.81±1.89μmol/L.化合物4和化合物10表现出对MV4-11、HCT116、HGC-27细胞株不同程度的抑制活性,化合物4对3种细胞株的IC50分别为22.87±2.41、29.58±1.96、48.85±2.05μmol/L,化合物10对3种细胞株的IC50分别为4.94±1.17、3.81±1.62、10.06±1.49μmol/L.本研究分离得到的单体化合物主要为甾类与环二肽类,其中化合物1-9,13-15,17-22从该菌种中首次分离得到,丰富了镰刀属真菌次生代谢产物的化学结构的多样性,可以进一步从生物间化学行为的角度探究川芎与其内生菌的相互作用规律和机制.This study examined the secondary metabolites of the endophytic fungus Fusarium proliferatum from Ligusticum chuanxiong Hort.and their cytotoxicity against three tumor cell lines.Twenty-three compounds were isolated and purified from the ethyl acetate extract of the fermentation broth via column chromatography,medium-pressure reversed-phase silica gel column chromatography,preparative HPLC,and other methods.Their structures were elucidated based on comprehensive spectroscopic analyses.The compounds were identified as follows:7-oxo-β-Sitosterol(1),7-oxo-β-stigmasterol(2),ergosterol peroxide(3),9,11-dehydroergosterol peroxide(4),5α,6α-epoxy-3β-hydroxyergosta-22-ene-7-one(5),5α,6α-epoxy-3β,8β,14α-Trihydroxy-5α-ergost-22E-en-7-one(6),3β-hydroxy-(22E,24R)-ergosta-5,8,22-trien-7-one(7),(4S,17R)-4-hydroxy-17-methylincisterol(8),(3β,5α,6β,22E)-ergosta-7,22-diene-3,5,6-triol(9),beauvericin(10),4-hydroxybenzaldehyde(11),cyclo-(L-prolyl-L-glycine)(12),cyclo-(Gly-D-Phe)(13),cyclo-(2-hydroxy-Pro-Gly)(14),cyclo-(L-Phe-Ala)(15),9-octadecenoic acid(Z)-,2’,3’-dihydroxypropyl ester(16),cyclo-(8α-hydroxy-Dprolyl-L-phenylalanine)(17),cyclo-(8β-hydroxy-L-prolyl-D-phenylalanine)(18),cyclo-(8α-hydroxy-L-prolyl-Lphenylalanine)(19),cyclo-(L-Pro-L-Leu)(20),cyclo-(L-Pro-D-Phe)(21),cyclo-(L-Pro-L-Phe)(22),and bassiatin(23).The MTT method was used to evaluate the cytotoxicity of compounds 1–7 and 10 against the MV4-11,HCT116,and HGC-27 cell lines.The results showed that compounds 3 and 5 demonstrated inhibitory activity against the MV4-11 cell lines,with IC50 values of 31.73±2.08 and 14.81±1.89μmol/L,respectively.Compounds 4 and 10 showed significant inhibitory activity against the MV4-11,HCT116,and HGC-27 cell lines;the IC50 values of compound 4 were 22.87±2.41,29.58±1.96 and 48.85±2.05μmol/L,respectively,while those for compound 10 were 4.94±1.17,3.81±1.62 and 10.06±1.49μmol/L,respectively.The compounds isolated were mainly steroids and cyclic dipeptides.Compounds 1–9,13–15,and 17–22 were isolated f

关 键 词：川芎 内生菌 Fusarium proliferatum 化学成分 细胞毒活性 

分 类 号：R284.1[医药卫生—中药学]