Study On The Extraction And Purification Of Nucleoside Components In Fresh Cordyceps Sinensis And Neuroprotective Effect

Parkinson's disease(PD)is a neurodegenerative disease that is second only to Alzheimer's disease.It is mainly caused by the loss of dopaminergic neurons in the substantia nigra of the midbrain,resulting in the inability of neurotransmitters to secrete normally.Dyskinesia,bradykinesia,etc.In addition,glial cell activation in the striatum is one of the early manifestations of PD,so behavioral tests,astrocyte and microglia number detection are also the main methods for judging Parkinson's disease.The current clinical drug for treating PD,dopamine hydrochloride,can only delay the onset of PD,and cannot prevent the progressive loss of dopaminergic neurons.Therefore,PD drug therapy and neuroprotective adjuvant therapy are also a good choice.Nucleoside components in food,such as creatinine,guanosine,and adenosine,have a certain degree of neuroprotection.The new resource food Cordyceps militaris is also rich in nucleoside components.Whether these nucleoside components have a certain auxiliary effect on PD by virtue of their neuroprotective effect is worthy of in-depth study.To this end,this study first compared the types and contents of nucleoside substances and other active ingredients in Cordyceps militaris from four production areas and three post-harvest processing methods,and selected better quality Cordyceps for extraction and purification of nucleoside components;PD mice were compared with the improvement of exercise ability,oxidative stress,neurotransmitters,etc.,and gradually analyzed the single nucleoside components(cordycepsin,adenosine,HEA),representative nucleoside composition(cordycepsin + adenosine + HEA),The neuroprotective effect of Cordyceps nucleoside extracts on PD and their interrelationships provide scientific basis for the auxiliary functions of Cordyceps in neurodegenerative diseases.The specific research results are as follows:1)Compare the content and types of active ingredients such as nucleoside substances in Cordyceps militaris in Yunnan,Shandong,Anhui,and Hubei,and use this to determine the better origin and processing method of Cordyceps militaris.By comparison,Cordyceps militaris in the four producing areas showed no significant differences in the types of nucleoside substances.Compared with other producing areas,Cordyceps militaris from Yunnan has more advantages in the content of active ingredients.Among them,ergosterol,cordycepic acid content and SOD enzyme activity are higher than other producing areas.)-Adenosine content is 0.19 mg / g,0.21 mg / g,0.24 mg / g.The order of the content of active ingredients in different processing methods is fresh> freeze-dried> dried.2)Compare the advantages and disadvantages of HPD100,NKA-? and D151 macroporous adsorption resins,and finally select NKA-? macroporous adsorption resin as the purification resin.By optimizing various adsorption processes of NKA-? macroporous adsorption resin,the best purification process is obtained: the p H of the sample solution is 8.0,the p H of the eluent is 7.0,the concentration of the eluent ethanol is 60%,and the sample volume is 50 m L,the flow rate of the eluent is 3 BV / h,the amount of eluent is 175 m L,and the purification frequency is 3 times.After purification,the samples were analyzed by liquid quality and the main nucleoside substances were cytidine,guanosine,adenosine,cordycepin,and HEA,and the purity was improved after three purifications.After three purifications,the purity of adenosine,cordycepin,and HEA in the purified fresh Cordyceps nucleoside were: 4.7%,14.8%,10.8%,freeze-dried: 9.8%,12.7%,9.4%,drying: 6.3 %,17.3%,5.2%.3)The effects of fresh and dried Cordyceps militaris purified products and single and mixtures of adenosine,cordycepin,and HEA on the behavior,neurotransmitter,glial cell morphology and number of Parkinson mice were studied.It was found that the fresh(dried)Cordyceps militaris nucleoside purified products,the three nucleoside single components and mixtures all have an improvement or relief effect on PD mice,among which,the fresh low dose is better than the drying low dose.The dose is better than the medium dose of adenosine and HEA,especially in terms of neurotransmitter content and number of glial cells,this difference is extremely significant.