| Fissistigma oldhamii var.longistipitatum is a variety of the genus Fissistigma(Annonaceae),which is widely distributed in the province of Zhejiang,Guangxi,Guangdong,Yunnan,Fujian,Taiwan etc.The genus of plants has wide range of pharmacological effects,such as cardiovascular protection,anti-gout,anti-tumor,anti-inflammatory and immunosuppressive effects,showing tremendous medicinal value.But as far as we know,its chemical constituents and biological activity has not been studied until now.Herein,we have conducted a series of researches on its chemical composition,biological activity and so on,taking Fissistigma oldhamii var.longistipitatum as the research object.The dried rhizomes of Fissistigma oldhamii var.longistipitatum were extracted by heating under reflux with 95%(v/v)aqueous ethanol.After removal of Et OH and water under reduced pressure,the crude extract was obtained and then mixed with double distilled water,which was partitioned successively with petroleum ether,dichloromethane,Et OAc and n-Bu OH to obtain four parts.The chemical constituents from the dichloromethane and Et OAc parts of the alcohol extract of the plants were separated and purified by silica gel,Sephadex LH-20,ODS,and semi-preparative HPLC.There are 28 compounds were identified primarily on the basis of references and spectroscopic analysis,such as UV,NMR,and HR-ESI-MS.All of these compoundsinclude22alkaloids,6flavonoids:(R)-1,2-methylenedioxy-3,9-dimethoxy-11-hydroxy-N-carbamoyl-noraporphine(1),3,10,11-trimethoxy-1,2-methylenedioxy-7-oxoaporphine(2),4’,5’-dimethoxy-2’-hydroxy-3’,6’-quinodihydrochalcone(3),fissistigamide A(4),fissistigamide B(5),liriodenine(6),oxocrebanine(7),oxobuxifolina(8),Oxocalycinine(9),fissistigmine(10),lysicamine(11),O-methylmoscatoline(12),atheroline(13),4,5-Dioxodehydroasimilobine(14),Norcepharadione B(15),piperolactam A(16),piperolactam C(17),Aristololactam AIa(18),Aristololactam Allla(19),Oldhamactam(20),goniothalactam(21),goniopedaline(22),aristolactam BIII(23),5,6,7,8-tetramethoxyflavone(24),5,8-dihydroxy-6,7-dimethoxyflavone(25),6-Hydroxy-5,7,8-trimethoxyflavone(26),7-Hydroxy-5,6,8-trimethoxyflavone(27),Methylpedicin(28).Compounds 1,2,3are new compounds and 8,26,27,28 were isolated from this genus for the first time,and all the above compounds were first reported from the plant.Application of UPLC-Q-TOF-MS/MS technology to rapidly analyze chemical constituents of the dichloromethane extract of Fissistigma oldhamii var.longistipitatum,according to the molecular ion peaks and MS~2 characteristic fragment ions,combined with the spectrum information of reference substances,fragmentation law of mass spectrum and related literatures,65 chemical constituents were preliminarily identified from the dichloromethane extract of Fissistigma oldhamii var.longistipitatum,including 44 alkaloids and 21 flavonoids.Taking 29 isolated monomeric compounds as the research object,the targets and signal pathways of Fissistigma oldhamii var.longistipitatum in the treatment of gout was predicted through network pharmacology to explore the potential mechanism of action.553 compound-related targets were obtained through the Swiss ADME and SEA databases,and 717 gout-related targets were retrieved from the Gene Cards database.Finally,113 potential related targets in the treatment of gout diseases by Fissistigma oldhamii var.longistipitatum were obtained.245 cell biological process(BP)items,34 cell composition(CC)items,and 49 molecular function(MF)items were screened by GO analysis and 56 signal pathways were screened by KEGG pathway enrichment analysis.The effective ingredients of Fissistigma oldhamii var.longistipitatum,6-Hydroxy-5,7,8-trimethoxyflavone,5,8-dihydroxy-6,7-dimethoxyflavone,goniothalactam,5,6,7,8-tetramethoxyflavone,lysicamine,etc.may act on GAPDH,IL6,IL2,ESR1,TLR4,XOD,VEGFA,MAPK1 and other key targets exert anti-gout effects by regulating TNF signaling pathway,Toll-like receptor signaling pathway,PI3K-Akt signaling pathway,IBD,NF-κB signaling pathway,etc. |
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