Study On Material Basis And Liver Protection Mechanism Of Ligustrum Lucidum Oil

Object:The purpose of this study was to clarify the chemical basis of Ligustrum lucidum oil and to explore the protective effect and mechanism of Ligustrum lucidum oil on chemical liver injury.Methods:1.The fingerprint of Ligustrum lucidum oil was constructed based on UPLC technology,and the chemical composition of Ligustrum lucidum oil was qualitative studied by LC-MS and GC-MS technology;2.Rat model of CCl₄-induced liver injury was used to observe the activities of AST and ALT in serum and the histopathological changes of liver,and to evaluate the pharmacodynamic effects of Ligustrum lucidum oil on chemical liver injury;3.The chemical components of Ligustrum lucidum oil confirmed by LC-MS and GC-MS were used as the database of chemical component,and the component-target network was constructed by network pharmacology technology and network analysis was conducted to predict the potential active components and targets of Ligustrum lucidum oil in improving CCl₄-induced liver injury in rats.Combined with KEGG pathway enrichment analysis to predict the signaling pathways related to Ligustrum lucidum oil improving CCl₄-induced liver injury in rats,the molecular mechanism of Ligustrum lucidum oil improving CCl₄-induced liver injury in rats was preliminarily explored;4.The plasma metabolite profiles of rats in control group,model group and Ligustrum lucidum oil administration group were constructed based on UPLC-Q-TOF-MS technology.PCA and PLS-DA analysis were used to characterize the differences of plasma metabolite profiles in each group.According to VIP>1 and P<0.05,biomarkers for liver injury induced by CCl₄ and liver protection by Ligustrum lucidum oil were screened.The results of metabolic pathway enrichment analysis and bionetwork analysis revealed the regulatory mechanism of Ligustrum lucidum oil on CCl₄-induced liver injury in rats.Results:1.49 chemical constituents of Ligustrum lucidum oil were identified by LC-MS,including 19 triterpenoids,21 lipids,5 flavonoids and 4 other components.19 volatile components were identified by GC-MS.;2.The results of the protective effect of Ligustrum lucidum oil on CCl₄-induced liver injury in rats showed that the serum AST and ALT activities of rats treated with 2%CCl₄ increased significantly（P<0.01）,and liver steatosis and cell edema,cytoplasm boundary blurred,accompanied by inflammatory cell infiltration and other damage.The activities of AST（P<0.01）and ALT（P<0.05）in serum of CCl₄-induced liver injury rats were significantly improved after the intervention of Ligustrum lucidum oil,indicating the protective effect of Ligustrum lucidum oil on chemical liver injury;3.Network pharmacology studies showed that oleanolic acid,ursolic acid,tormentic acid,linoleic acid and other components in Ligustrum lucidum oil act on PPARγ,PPARα,TNF and MAPK3,reduce inflammation and regulate apoptosis by regulating PI3K-Akt,TNF and MAPK signaling pathways,and jointly play a role in liver protection;4.In plasma untargeted metabolome analysis,PCA and PLS-DA analysis were used to screen 27 differential metabolites of CCl₄-induced liver injury,including L-valine,L-leucine andγ-linolenic acid,which were closely related to amino acid metabolism and unsaturated fatty acid biosynthesis.The 20 metabolites of L-valine,L-leucine andγ-linolenic acid were significantly reduced after the intervention of Ligustrum lucidum oil（P<0.01）.Through the enrichment of metabolic pathway and biological network analysis,linoleic acid,palmitic acid,tormentic acid,oleanolic acid,ursolic acid and other components of Ligustrum lucidum oil can inhibit the activity of PLA2,improve the metabolism of fatty acids and amino acid metabolism in mice to reduce inflammation,restrain cell autophagy and necrosis,which played a protect liver function.Conclusions:In this study,it was found that the Ligustrum lucidum oil mainly contains triterpenoids,oils,volatiles and other components,which have good protective effect on liver injury induced by CCl₄ in rats.Oleanolic acid,ursolic acid and tormentic acid in Ligustrum lucidum oil can reduce liver inflammation by inhibiting the activity of PPARs and PLA2.Moreover,TNF and MAPK signaling pathways can inhibit cell necrotic apoptosis,and thus play a role in liver protection.