Identification Of Triterpene Acid Components In Radix Glehniae And Study On Its Liver Protection Mechanis

Triterpenic acids compounds are an important class of traditional Chinese medicine chemical components of medicinal and edible plants,which have biological activities such as liver protection,immune regulation,lowering blood glucose and lipids.Lycopus lucidu Turcz.is the root part of the traditional Chinese medicine Lycopus Lucidu,which is widely distributed in most areas of China.It is a perennial herb.Because of its rich nutrition and tenacious vitality,it is loved by people,and it is a good homologous plant resource of medicine and food.Using L.lucidu as experimental raw material,three extraction methods including ultrasound-assisted extraction method,heated method and reflux extraction method were used to extract triterpenic from L.lucidu.Three triterpenic acids(betulinic acid,oleanolic acid,ursolic acid)in L.lucidu were detected by HPLC.Response surface was used to optimize the extraction process of triterpenic acids from L.lucidu and its content was determined.The total triterpenic acids was determined by UV method and purified by macroporous resin.The purification process parameters were optimized.The network pharmacology and molecular docking techniques were established to further explore the hepatoprotective mechanism of the triterpenic acids components of L.lucidu.The main research results are as follows:(1)Orthogonal experiment was designed on the basis of single factor experiment,three methods of ultrasonic-assisted extraction,heating extraction and reflux extraction were selected,and the total contents of three triterpenic acids(betulinic acid,oleanolic acid and ursolic acid)in L.lucidu were taken as the index to optimize the optimal technological parameters of each extraction method.The contents of triterpenic acid in L.lucidu were ranked from high to low as: betulinic acid>ursolic acid>oleanolic acid.(2)Taking the total contents of three main triterpenic components in L.lucidu betulinic acid,oleanolic acid and ursolic acid as evaluation index,four factors including solvent concentration,extraction time,extraction temperature and solid-liquid ratio were selected on the basis of single factor experiment.The effects of four factors on the extraction effect of triterpenic acids of L.lucidu were ranked as follows: ethanol volume fraction > materialliquid ratio > extraction temperature > extraction time,the ethanol volume fraction had significant effects(P<0.01),solid-liquid ratio and extraction temperature had significant effects(P<0.05),and the interaction between the two factors had no significant effects on the extraction of L.lucidu triterpenic acids(P>0.05).(3)The total triterpenic acids of L.lucidu was adsorbed by macroporous resin,and the total triterpenic acids of L.lucidu was easily adsorbed by AB-8 resin by comparing five macroporous resins with adsorption rate and resolution rate.The purification effect of triterpenic acids by four single factors was as follows: elution concentration > sample concentration > loading speed > eluent velocity.The highest recovery rate of total triterpenic acids from L.lucidu increased from 60.08% to 67.36%.(4)Triterpenic acids compounds and their targets were obtained by searching TCMSP,Pharm Mapper databases,and the Target was obtained by Swiss Target Prediction and uniprot databases.Liver disease targets were obtained from OMIM and Genecards databases,and common targets were obtained from Venny2.1 online software mapping tool platform.Using Cytoscape 3.9.1 software,construct the composition-Target-disease network and enter it into the STRING database to retrieve the PPI network of construction protein interactions.GO and KEGG enrichment analysis was performed on the David database.The docking result was verified by Molecular Docking.In conclusion,according to the operability and simplicity of the experiment,the heating extraction method was selected to optimize the extraction and purification process of triterpenic acids from L.lucidu.The hepatoprotective mechanism of triterpenic acids compounds from L.lucidu was explored based on network pharmacology and molecular docking technology.