Study On Chemical Constituents And Biological Activities Of Chroogomphus Rutilus (Schaeff.) O.k. Mill

Objective: To identify the main active components of anti-hepatoma effect of Chroogomphus rutilus based on spectrum-effect relationship and component knock-out technology,and to study the network pharmacology of the main active components of Chroogomphus rutilus to predict the mechanism of action.Method:1.Firstly,MTT method was used to investigate the inhibitory effect of different polar parts of Chroogomphus rutilus on HepG2 cells,and the best active part was selected;2.HPLC-DAD was used to establish the chemical fingerprints of Chroogomphus rutilus from different areas,and the anti-hepatoma activity of each sample was analyzed by PLS with SIMCA software;3.HPLC method was used to knock-out the active components which were screened in the early stage,and LC-ESI-MS/MS was used to speculate the possible chemical structure.MTT method was used to evaluate its anti-hepatoma activity and identify the main active components in Chroogomphus rutilus;4.The network pharmacology method was used to predict the anti-hepatoma mechanism of the main active components in Chroogomphus rutilus.Result:1.In addition,the IC50 value of dichloromethane was significantly lower than that of other drug groups,indicating that the main active components of Chroogomphus rutilus in anti hepatocarcinoma were in dichloromethane layer;2.The inhibitory effect of Chroogomphus rutilus dichloromethane extract from different places on HepG2 cells was different.When the sample concentration changed,the inhibitory effect on HepG2 cells was also different.Through the regression coefficient and VIP value of PLS,peaks 1,3,5 and 13 were found to be the compounds with positive correlation and strong correlation with anti-hepatoma activity.3.Peak 3(Acetic acid 2R-(4,8-dimethylnona-3,7-dienyl)-8-hydroxy-2-methyl-2Hchromen-6-yl ester),5(Ergosta-7,22-dien-5,6-epoxy-3-ol)and 13(Ergosterol)inhibited the proliferation of HepG2 cells,which was consistent with the spectral effect,of which peak 13 had the strongest inhibition,but peak 1(Palmitic monoglycerol ester)knock-out components promoted the proliferation of HepG2 cells.4.Using Pharm Mapper server to predict the target of ergosterol,224 potential targets were obtained.In the OncoDB.HCC database,378 genes related to liver cancer were collected.By mapping and comparing the target of Ergosterol with the target of liver cancer related genes,36 potential target genes were screened out.The analysis of the network interaction network of ergosterol on the target protein of liver cancer showed that there were a lot of interactions among the targets,among which MAPK1,SRC,CASP3,HSP90AA1 and EGFR were more important Protein target.The mechanism may be related to E-cadherin signaling events,Alpha6Beta4 Integrin,Nongenotropic Androgen signaling,ErbB receptor signaling network,TNF receptor signaling pathwadeng and other pathways.Conclusion: On the basis of spectral effect relationship,the main active components of anti-hepatoma in Chroogomphus rutilus were identified by component knock-out technology.At the same time,the anti-hepatoma mechanism of ergosterol,one of the main active components in Chroogomphus rutilus,was clarified preliminarily,which provided scientific basis for the research of pharmacodynamic substance basis,medicinal and food development of Chroogomphus rutilus.