Material Foundation Research Of Antitumor Effective Parts Of Capparis Spinosa Based On Network Pharmacology

Objective:To study the active ingredients and antitumor activity of different polar parts of capers.The anti-tumor active ingredients and mechanism of action of chloroform,petroleum ether and ethyl acetate sites were predicted,and the prediction results of ethyl acetate site were confirmed by in vitro experiments.Methods:The the crude powder of Capparis fruits were taken and extracted with 85% ethanol to obtain the crude ethanol extract of Capparis capsicum,and the different effective parts including petroleum ether,chloroform,ethyl acetate,and water-soluble parts were extracted separately,cck-8 method was used to determine the antitumor activity of each part in vitro;Column chromatography was used to separate the chemical constituents of effective parts of petroleum ether from Capparis fruits.Physical and chemical properties and spectral data were used to analyze the chemical structure;The anti-tumor active ingredients and mechanism of action of chloroform,petroleum ether,and ethyl acetate were predicted by network pharmacology and the prediction results of the ethyl acetate site were verified by in vitro experiments.Results:The polar extracts such as total alcohol extract,chloroform fraction,petroleum ether fraction,ethyl acetate fraction,and water extract of capparis fruit were obtained by extraction,the cck-8 measurement results show that each polar site has inhibitory effects on different tumor cells in time-and concentration-dependent way;Three compounds were separated and identified by physical and chemical means and NMR and other spectral techniques from the petroleum ether site by column chromatography.They were 2-Ethyl ethyl-1-tetradecene,β-sitosterol,and carotene;According to the prediction of network pharmacology,from the chloroform components of Capparis fruit,Wogonin and Oroxylin A were selected as representative active ingredients,and PI3K-Akt signaling pathway and nitrogen metabolism pathway were selected as representative signaling pathways;From the volatile constituents of petroleum ether in Capparis fruits,arachidic acid,n-docosadecane,methyltocols,monoacetin,ethyl stearate and monopalmitate were elected as representative active ingredients,and nitrogen metabolism was selected as representative signaling pathways;From the components of ethyl acetate in Capparis fruits,isorhamnetin,kaempferol and apigenin were selected as representative active ingredients,and PI3K-Akt signaling pathway and nitrogen metabolism were selected as representative signaling pathways;In verification test,cck-8 results showed that isorhamnetin,kaempferol,and apigenin all significantly inhibited the proliferation of human liver cancer cells HepG-2 in a time-and dose-dependent manner;Flow cytometry results showed that three representative active ingredients acting on HepG-2 cells for 48 hours could increase the apoptosis rate of the cells,and the effects were dose-dependent,and they all had certain effects on cell cycle;Western blotting results show:Compared with the control group,the expression of Akt and PI3K protein in each dose group of the experimental group was reduced,indicating that the ethyl acetate site can block the PI3K/Akt pathway.Conclusion:This study found that Capparis fruits are rich in various active ingredients and have multiple pharmacological effects.In this study network pharmacological method was applied to conduct a preliminary study on the antitumor mechanism of active parts of Capparis fruits,and the potential antitumor mechanism of capers fruit extract was clarified from the perspective of analysis,which laid the foundation for the later research of capers.