Chemical Constituents From The Aerial Parts Of Cirsium Japonicum And Their Activities

Cirsium japonicum,a perennial herb belonging to the genus Asteraceae,has been used as a kind of Chinese traditional medicine to stop the bleeding over a thousand years.In order to screen novel botanical fungicides,the investigation on the chemical constituents and its activities of the aerial parts of Cirsium japonicum was carried out.The plant was extracted with methanol.After liquid-liquid extraction,separation by silica gel and Sephadex LH-20 column chromatography,13 compounds were finally obtained from the ethyl acetate phase.By comparing the NMR and MS data of the compounds with the data in literatures,they were identified as ciryneol D（1）,12β,20（S）-dihydroxydammar-24-en-3-one（2）,9（11）,12-dieneoleana-3β-ol（3）,taraxasterol（4）,3β-hydroxystigmast-5-en-7-one（5）,3β-hydroxyl-12（13）-en-olean-11-one（6）,p-coumaric acid（7）,obtucarbamate A（8）,Methyl 4-hydroxybenzoate（9）,4,4’-diphenylmethane-bis（methyl）carbamate（10）,quercitrin（11）,kaempferol-3-O-α-L-rhamnoside（12）,and chrysoeriol-7-O-2’-O-6’-O-acetyl-β-D-glucopyranosyl-β-D-glucopyranoside（13）.Among them,2,3,6,8,10,12 and 13 were isolated from this plant for the first time.Ciryneol D（1）showed inhibitory effect against the mycelia growth of five species of plant pathogenic fungi.Among all the tested microorganisms,1 showed best effect against Valsa mali with the EC₅₀0 of 23.21μg/mL.Ciryneol D also exhibited significant inhibition on the spore germination of Curvularia lunata,Colletotrichum gloesporioides and Botrytis cinerea,with the EC50 values of 27.52,26.41 and 81.93μg/mL,respectively.Besides,Bacillus cereus was sensitive to ciryneol D with the MIC of 50μg/mL.In addition,ciryneol D can inhibited canker lesion expansion of Valsa mal on excised twigs significantly,with the EC₅₀ of 32.82μg/mL.Establish quantitative analytical method to compare the content of ciryneol D in the aerial parts and roots of Cirsium japonicum.HPLC method with Hypersil BDS C18 column（4.6 mm×250 mm,5μm）,and the mixture of Methanol-H2O（75:25）as the mobile phase was adopted.The flow rate was 0.6 mL·min^-1,and the UV detection wavelength was 245 nm.The assay displayed that the content of ciryneol D was richer in roots.In terms of the aerial parts of Cirsium japonicum useless in medicinal value,they can be a kind of source for the development of botanical fungicides.