Studies On Chemical Constitutions Of Ganoderma Gurtisii And Inhibitory Effect Of Triterpenoids On NO Released By LPS-induced Microglia

Objective:To study the chemical constituents of the fraction with anti-inflammatory activity from Ganoderma curtisii(Berk.)Murrill and determine anti-inflammatory activity of ganoderma triterpenoids.This work will provide the fundamental data for the protection of neuroinflammation.Methods:The chemical constituents of the 85%ethanol extract of fruting bodies of Ganoderma curtisii.were studied by means of various chromatographic techniques such as silica gel,reversed phase ODS column,Sephadex LH-20 and PHPLC chromatography.Their chemical structures were deduced on the basis of physicochemical property and spectroscopic analysis.The model of inflammation was established by inducing BV-2 microglia cell with lipopolysaccharide(LPS)leading to nitric oxide(NO)production,the cell viability on method was evaluted by MTT assay,the concentration of NO2-into the supematant was determined by Griess method.The anti-inflammatory activity of Ganoderma triterpenoids was measured with the above model.Results:The EtOAc extract of dried chipped fruiting bodies of Ganoderma curtisii was subjected to extensive column chromatography to afford 34 compounds.Their structures were identified as follows:3?,12?-dihydroxy-7,11,15,23-tetraoxo-lanost-8,20-dien-26-oic acid(1),15a-hydroxy-3,11,23-trioxo-lanost-8,20-dien-26-oic acid(2),12?-acetoxy-.3,7,11,15,23-pentaoxo-lanost-8,20-dien-26-oic acid(3),3,7,12-trihydroxy-1 1,15,23-trioxo-lanost-8,20-dien-26-oic acid(4),3,7,15-trihydroxy-4-(hydroxymethyl)-11,23-dioxo-lanost-8-en-26-oic acid(5),ganodermadiol B(6),ganoderic acid C2(7),ganodermatriol(8),ganodermanontriol(9),methyl lucidenate L(10),ganoderic acid C6(11),ganoderic acid G(12),ganoderic acid B(13),ganoderic acid D(14),ganoderenic acid D(15),ganoderic acid A(16),7?,12?-dihydroxy-3,11,15,23-tetraoxo-5a-lanost-8-en-26-oic acid(17),ganoderic acid H(18),ganoderenic acid A(19),ganoderic acid E(20),ganolucidic acid A(21),ganoderic acid J(22),methyl ganoderate A(23),3?,15?-dihydroxy-7,11,23-trioxo-lanost-8-dien-26-oic acid(24),ganoderic acid F(25),ganoderenic acid K(26),20-Hydroxylganodric acid G(27),12?-acetoxy-3?-hydroxy-7,11,15,23-tetraoxo-lanost-8,20E-diene-26-oic acid(28),ganoderic acid K(29),ergosterol(30),Ergosta-7,22-dien-3?-ol(31),Ergosterol peroxide(31),6,9-epidioxyergosta-7,22-dien-3?-ol(33),liquiritigenin(34).Among them,there were 29 triterpenoids,three steroid and a flavonoid.Compound 1 was new compound,compounds 2?5 were obtained from ganoderma for the first time.The inhibition abilities of 27 compounds against NO production on LPS-induced BV-2 microglia have been examined.The results showed that 26 Ganoderma triterpenoids had no effect on the survival rate of BV-2 microglia cell,which better suppressed NO production induced by LPS in BV-2 microglia.Compound 4,5,12,17,19,20,23,24,29 showed potent inhibitory activity on NO production in BV-2 cells stimulated by LPS,and their IC50 values were less than 10?M.Conclusion:29 triterpenoids were isolated from the EtOAc extract of Ganoderma curtisii,among them,9 compounds showed the strong inhibition on the NO releasing.Thus,triterpenoids are presumed the active components with anti-imflammotary and perspective of anti-neuroinflammation.