Screening, Isolation, Structure Elucidation Of Medicinal Fungi's Neuroactive Components

Alzheimer's diease is a kind of neuro-degenerative diease mainly affecting both cognition and behavior with unkown etiology.PC12 cell is a immature neuron cell line sources of rat adrenal pheochromocytoma cells,has been widely used in pathological research and drugs screening of nervous systerm diease.Medicinal fungi has been used to treat various human ailments for over 2,000 years in China.Medicinal fungi contain a lot of bioactive components,such as polysaccharide,terpenes,flavonoids and so on,have been reported to exhibit various pharmaceutical effects including regulating immune,antitumor, antibiotic,neurotrophin etc.At present,some medicinal fungi have been reported to exhibit neurotrophin and protecting effect of neuron damage,but previous studies focused mainly on promoting PC12 cell differentiation.Studying on tracking on neuroactive components of medicinal fungi's isolation has never been reported.In this study,promoting PC12 cell neurite outgrowth model and PC12 cell oxidative-stress damage model were built.Extracts of Four medicinal fungi was screened by PC12 models,and isolation of Hericium erinaceus lipophilic extracts was tracked on antioxidative-damage active components in PC12 cell.Reserch shows that,some Ganoderma lucidum stains and Hericium erinaceus lipophilic extracts have a certain function to PC12 cell neurite outgrowth,but the effect is not obvious.Lipophilic extracts of four species medicinal fungi play a certain roles in protecting effect against oxidative-stress damage in PC12 cell.In each medicinal fungus,the best neurotive site including:GC15 stains' CVR is(88.36±1.8)%at the concentration of 50μg/mL;DT4's eve.is(68.23±0.3)%at the concentration of 50μg/mL;HW's CVR is (78.08±3.1)%at the concentration of 50μg/mL;FC5's CVR is(81.2±3.1) at the concentration of 150μg/mL.The most fractions isolated from Hericium erinaceus have a certain protecting effect.HE at concentration between 5μg/mL and150μg/mL promoted cell viability in a dose-dependent manner,CVR of each concentration is(40.35±1.1)%,(63.53±4.1)%, (69.26±3.3)%.HE5-1-3 isolated from HE promoted cell viability also in dose-dependent manner,CVR of each concentration is(42.54±3.2)%,(50.48±0.5)%,(62.33±1.3)%. The CVR of compounds HC3-7-1 is(46.37±6.1)%,(58.73±4.1)%,(61.44±2.7)%.The two compounds all posses a neurotive,but they are lower than the crude extracts.According to the polarity,Hericium erinaceus lipophilic extracts were isolated with petroleum ether,chloroform,ethyl acetate,and BuOH respectively.Then,the fraction were purified using silica gel column chromatography by a gradient elution with different elution systerm.,two compounds was obtained.Analyse with TLC in positive and reversed phase, two compounds are single compound,their structures elucidated from ¹H-NMR,¹³C-NMR and MS spectral data.Compound HE5-1-3-1 is nicotinamide.