Isolation And Activities Evaluation Of Terpenoid Compounds Of Ganoderma Applanatum

Objective:To study chemical constituents of Ganoderma applanatum(Per.)Pat.,and determine the potential anti-inflammatory and neuroprotective effects of isolated compounds.This work will provide the foundamental material for illustrating the neuroprotection of G.applanatum.Methods:The chemical constituents of 85%EtOH extract of fruiting bodies of G.applanatum were studied using various chromatographic techniques.The structures of those compounds were characterized by spectroscopic analysis.Lipopolysaccharide(LPS)-induced BV-2 microglia cells was used to evaluate the anti-inflammatory effect of compound 1 by determing the concentration of NO,cytokines,and chemokines in supernatant,and the content of ROS in B V-2 cells.The conditioned medium of BV-2 cells was used to incubate with HT22 murine hippocampal neuronal cells,and to evaluate the neuroprotective effect of compound 1 by various staining methods and determining the level of MAP2 in HT22 cells.Results:Nine compounds were isolated from the fruiting bodies of Ganoderma applanatum,and their structures were identified as following:15?-hydroxy-7?,8?-epoxy-4,4,14?-trimethyl-3,12,20-trioxo-5?-pregn-9(11),16(17)-diene(1),1 5?,20?-dihydroxy-7?,8?-epoxy-3,12,23-trioxolanosta-9,16-dien-26-oic acid methyl ester(2),Ganoderenic acid E(3),Applanone E(4),Applanoic acid B(5),Gibbosic acid A(6),Lingzhiol(7),Ergosterol peroxide(8),Ergosta-7,22-dien-3-one(9).Among them,there were six highly oxygenated C24 or C30 lanostane triterpenoids with 7,8-epoxy group including three new compound,one meroterpenoid which was obtained from G.applanatum for the first time,and two known steroids.The anti-inflammatory and neuroprotective activities of compound 1 was evaluated.The results showed that compound 1 effective inhibited the release of NO,IL-1?,IL-6,TNF-?,GM-CSF,MCP-1 and MIP-2 on LPS-induced BV-2 cells with no cytotoxicity,and decreases the level of ROS in the cells.And compound 1 could decrease apoptosis cell populations and increase the level of MAP2 to achieve neuroproctive effect.Conclusion:The triterpenoids from G.applanatum are different from other fungus of Ganoderma.This research found an active triterpenoid possessing neuroprotective effect by inhibiting over-activated microglia cells.This work helps further research to find more bioactive compounds,and can elucidate the connection of neuroprotection effect and chemical compositions of G.applanatum.