Chemical Components And Preliminary Screening Of Anti-inflammatory Activity Analysis Of Ganoderma Lucidum

Objective:Ganoderma has been used as a traditional Chinese medicinal material for more than 2000 years in China.The earliest record of Ganoderma was in the“Shen Nong’s Materia Medica”and Ganoderma was known as immortal grass.For a long time,the research on components of Ganoderma mainly focuses on triterpenoids and polysaccharides,which explains the source of partial efficacy of Ganoderma.But the pharmacological action of Ganoderma is very extensive,its pharmacodynamic material basis remains to be further clarified.Therefore,in order to further explore the chemical composition of Ganoderma and reveal the basis of its pharmacodynamic material.The chemical composition of Ganoderma Lucidum were isolated and identified,and new compounds were preliminarily screened for anti-inflammatory activity.Methods:The powders of fruiting bodies of Ganoderma Lucidum（500.0 kg）were extracted with 80%Et OH/aqueous solution（2×3000 L×2-3 h）to yield a crude extract（24.6 kg）.The crude extract was extracted with ethyl acetate:water=5:4 for 8 times followed by removing solvents to afford 11.5 kg extracts.The Et OAc extract was separated via column chromatography（MCI gel CHP 20P,Sephadex LH-20,silica gel,RP-18）,PTLC and HPLC to obtain monomer compound.The structures of compounds were identified by spectroscopic methods(¹H-NMR,¹³C-NMR,DEPT,COSY,HSQC,HMBC,MS).Finally,the cytotoxicity of the new compounds were detected by CCK-8.The effect of the new compounds on the expression of inflammatory factors in RAW264.7 were detected by fluorescence quantitative PCR.Results:20 compounds including 15 new compounds（5 steroids,1 triterpenoid and 9 meroterpenoids combine with triterpenoids）and 5 know compounds were isolated from Ganoderma Lucidum.New compounds were assayed for cytotoxicity and preliminarily anti-inflammatory activity.The IC₅₀ of compound 2,3,5,6,7,8,9,10,13,15,16,17,18,19,20 were 24.56μM,20.03μM,29.86μM,33.24μM,18.35μM,more than 100μM,23.02μM,21.23μM,26.52μM,77.48μM,25.61μM,45.73μM,56.16μM,46.25μM,30.57μM,respectively.Compounds 16 and 19 both significantly inhibited the m RNA expression of IL-1βand IL-6 at the concentration of 10μM.At the concentration of 10μM,compound 18 has statistically different inhibitory effects on m RNA expression of IL-1β,which can significantly inhibit the m RNA expression of IL-6.Conclusion:In this study,15 new compounds were isolated from Ganoderma Lucidum.Compounds 16,18 and 19 showed anti-inflammatory potential.This study enriched the chemical component and discovered new medicinal substances of Ganoderma,which laid a foundation for the subsequent study of the biosynthesis and pharmacodynamic mechanism of meroterpenoids combine with triterpenoids.