Study On Hepatoprotective Constituents From Acer Tegmentosum And Daphne Koreana In Changbai Mountain Area

Objective:To study the liver protective activities of Daphne koreana and Acer tegmentosum in Changbai Mountain area,screen the effective components and separate the active monomers,determine the cytotoxicity and liver protective effects of the two plants and predict the possible targets and pathways by using network pharmacology.Methods:The traditional Chinese medicine chemical component extraction method is used for reflux extraction of the medicinal materials of Acer tegmentosum and Daphne koreana to obtain an ethanol extract.Extracted with petroleum ether,dichloromethane,ethyl acetate and n-butanol respectively to obtain four extraction layer extracts.A variety of chromatographic separation techniques were used to separate the main chemical components and structural identification of the ethyl acetate layer of Acer tegmentosum and Daphne koreana.The growth inhibition of HeLa and HepG2 cells was determined by MTT method.The total extracts of Acer tegmentosum and Daphne koreana were selected to study the hepatoprotective effect of carbon tetrachloride-induced acute liver injury model in mice,and the ALT and AST indexes were detected to determine the hepatoprotective activity of the two plants.Based on the network pharmacology,the active ingredients of Acer tegmentosum,Daphne koreana and the targets of acute liver injury were predicted.Results:In the study of the chemical constituents of the ethyl acetate extract of Acer truncatum,4-hydroxyphenylethyl-1-O-β-D-[6-[6’-O-hydroxybenzoyl)]-glucopyranoside(1)was isolated.In the process of compound research on Daphne koreana,11 monomer compounds were isolated,and the structure was determined by nuclear magnetic resonance hydrogen spectroscopy and carbon spectroscopy to detemine the structure:β-sitosterol(2),apigenin(3),umbelliferone(4)daphnoretin(5),dibutyl phthalate(6),daphnetin(7),naringenin(8),chamaejasmin(9),daphnin(10),syringin(11),daphnodorin E(12).The results of liver protection activity tests of the total extracts of Acer iegmentosum and Daphne koreana showed that their ALT and AST indexes were lower than the model group,and the cytotoxicity was low.Acer tegmentosum ethyl acetate layer,Daphne koreana dichloromethane layer and Daphne koreana ethyl acetate layer group showed better anti-Taxoplasma gondii effect The network pharmacology results show that the components like daphnetin and umbelliferone in Acer tegmentosum and Daphne koreana are related to many targets of acute liver injury and can be used as potential drugs for treating acute liver injury;GAPDH,TP53,AKT1,ALB and MAPK3 are the key target in its protein interaction network;pathways related to acute liver injury include MAPK signaling,NF-κB signaling,JAK-STAT signaling,etc.Conclusion:In this experiment,A new compound was isolated from Acer tegmentosum,compound 6,9 and compound 12 were isolated from Daphne koreana for the first time.Both Acer tegmentosum and Daphne koreana showed certain hepatoprotective activity,their mechanism of action needs further study.