蜜环菌发酵液及其菌丝体的化学成分研究

Chemical constituents from Armillaria mellea fermentation broth and its mycelia

蜜环菌(Armillaria mellea)是药用植物天麻(Gastrodia elata)营养生长阶段的共生真菌。为研究蜜环菌的化学成分和生物活性,本实验综合运用MCI树脂、正相硅胶、Sephadex LH-20凝胶及半制备RP-HPLC等色谱分离技术对蜜环菌发酵液及其菌丝体的95%乙醇提取物进行系统分离,得到26个化合物。根据化合物的理化性质和波谱数据,分别鉴定为十四烷酸(1)、α-亚麻酸(2)、油酸(3)、亚油酸(4)、亚油酸乙酯(5)、8(R),11(S)-二羟基-9 Z,12 Z-十八烷二烯酸(6)、2-油酰甘油(7)、1-亚油酸甘油酯(8)、二羟丙基-油酸(9)、亚油酸甲酯(10)、甘油1,3-二亚油酸酯(11)、(5 Z,9 Z)-17-甲基-非十七烯酸-5,9-二酯(12)、(2 S)-α-(9′Z,12′Z,15′Z)-十八碳三烯酸(13)、麦角甾醇(14)、(22 E,24 S)-5α,8α-环二氧-6,22-麦角甾二烯-3β醇(15)、对羟基苄基乙基醚(16)、尿嘧啶(17)、环D-脯氨酸-D-异亮氨酸(18)、环D-脯氨酸-D-亮氨酸(19)、环D-脯氨酸-D-苯丙氨酸(20)、脯氨酸(21)、环D-脯氨酸-L-缬氨酸(22)、环D-脯氨酸-D-缬氨酸(23)、4-(2-羟乙基)-5-甲基噁唑(24)、环丙氨酸-缬氨酸(25)、环丙氨酸-脯氨酸(26)。其中,化合物5~13、16和18~26是首次从蜜环菌中分离获得。体外生物活性研究表明化合物14和15对LPS诱导的RAW 264.7细胞炎症反应具有抑制作用。

Armillaria mellea is a symbiotic fungus essential for the nutritional growth stage of the medicinal plant Gastrodia elata.To investigate the chemical constitutes and bioactivities of A.mellea,the 95%ethanol extract of A.mellea was repeatedly chromatographed over silica gel,MCI gel,Sephadex LH-20 and reversed-phase(RP)-HPLC,affording twenty-six pure compounds(1-26).Their structures were identified as tetradecanoic acid(1),α-linolenic acid(2),oleic acid(3),linoleic acid(4),ethyl linoleate(5),8(R),11(S)-dihydroxy-9 Z,12 Z-octadecadienoic acid(6),2-linoleoylglycerol(7),1-monolinolein(8),9-octadecenoic acid-2′,3′-dihydroxy propyl ester(9),methyl linoleate(10),glycerin 1,3-dilinoleate(11),(5 Z,9 Z)-17-methylnonadeca-5,9-dienoate(12),(2 S)-α-(9′Z,12′Z,15′Z)-octadecatrienoic acid(13),ergosta-5,7,22-triene-3β-ol(14),(22 E,24 S)-5α,8α-epidioxy-24-methyl-cholesta-6,9(11),22-trien-3β-ol(15),p-hydroxybenzylethyl ether(16),uracil(17),cyclo(D)-Pro-(D)-Ile(18),cyclo(D)-Pro-(D)-Leu(19),cyclo(D)-Pro-(D)-Phe(20),proline(21),cyclo(D)-Pro-(L)-Val(22),cyclo(D)-Pro-(D)-Val(23),4-(2-hydroxyethyl)-5-methyloxazole(24),cyclo-(Ala-Val)(25),cyclo-(Ala-Pro)(26),respectively,by comparision of their NMR data and optical rotation values with those reported in the literature.Among them,compounds 5-13,16,and 18-26 were obtained for the first time from A.mellea.In in vitro bioassay,compounds 14 and 15 showed moderate effects against LPS-induced inflammatory responses in RAW 264.7 cells.