Study On Chemical Constituents Of Penthorum Chinense Pursh And Treatment Of Alcoholic Liver Injury In Mice

Penthorum chinense Pursh is a local herbal medicine of the Miao people,which has the functions of liver protection,yellowing and detoxification.The Penthorum chinense Pursh is mainly grown in the southwest of China,and the producing area in Gullin County has the best quality.Penthorum chinense Pursh is rich in chemical components,including flavonoids and glycosides,lignans,organic acids,volatile oil,etc.Basic pharmaceutical and clinical studies have shown that in addition to the protective effect of liver damage,it also has anti-tumor,anti-oxidation,antiinflammatory and other pharmacological activities.In addition,tea bags and drinks made from Penthorum chinense Pursh have been widely sold in the market and can be eaten as vegetables.Because of its great value in medicinal and edible aspects and broad development prospects,the related research,development and utilization of Penthorum chinense Pursh has made some progress.However,there are still some problems,such as insufficient research on the activity of Penthorum chinense Pursh,no reports on the research and development of new products other than liver protection,the basis of drug efficacy is not clear,and there is no more strict quality standard.So far,Penthorum chinense Pursh has not been included in the "Chinese Pharmacopoeia",which limits the further development and utilization of Penthorum chinense Pursh.In this study,a series of experiments were conducted around Penthorum chinense Pursh driving,and the main methods and results were as follows:Methods: HPLC-QTOF/MS technique was used to analyze the chemical constituents of 70% ethanol extract from the leaves of Penthorum chinense Pursh,and the chromatographic peaks in the total flow pattern were identified by mass spectrometry ion fragments.The hepatoprotective activity of the water extract of Penthorum chinense Pursh was studied by establishing a mouse model of acute alcoholic liver injury,including HE staining of liver sections and analysis of serum TG,TC,AST and ALT contents.By sequencing 16 s RNA of colon contents of mice,the differences of intestinal flora species in blank group,alcohol model group and Penthorum chinense Pursh administration group were analyzed.The active ingredients,protein targets and action pathways involved in the treatment of alcoholic liver injury were analyzed by network pharmacology.Results:(1)38 components were identified from the LC-MS diagram of 70%ethanol extract from the leaves of Penthorum chinense Pursh.They were Chebulic acid,Gallic acid,Protocatechuic acid,Methyl gallate,catechin,Rutoside,Vanillic acid,Helicin,Ethylgallate,Syringaldehyde,Ellagic acid,Isoquercitrin,Quercetin-3-O-β-Dxylopyranoside,Cynaroside,kaempferol-3-O-Arabinoside,Apige nin-7-O-β-DGlucoside,Pinocembrin-7-O-β-D-Glucoside,pinocembrin-7-O-[4",6"-(S)-hexahydroxydiphenoyl]-β-D-glucose,pinocembrin-7-O-[3’’-O-galloyl-4’’,6’’-(S)-hex ahydroxydiphenoyl]-β-D-glucose,Quercetin-3-O-robinobioside,Kaempferol-3-O-rut inoside,Scopoletin,Quercetin-3-O-arabinoside,Quercitrin,luteolin,Baicalein-7-O-rha mnoside,Pinocembrinchalcone,Quercetin-3-O-sambubioside,Quercetin-3-O-robinosi de-7-O-glucoside,Quercetin-5,7-O-diglucoside,Xanthone C,Evanesin D,Thonningianin A,Evanesin C,Thonningianin B,2’,6’-dihydroxy-dihydrochalcone-4’-O-(3"-O-gallic)-β-D-glucopyranoside,2’,4’,6’-trihydroxy-dihydrochalcone-4’-β-Dglucoside,Loconin-7-O-glucoside,2,6-dihydroxyacetophenone-4-O-β-Dglucopyranoside.(2)HE staining results of liver sections showed that in the alcohol model group,there were a large number of liver cell necrosis,inflammatory infiltrating cells,volume expansion of liver cells,blurred boundaries between cells,irregular arrangement of liver cell cords,cavity structure in part of liver cytoplasm,and congestion and expansion of local hepatic sinuses and hepatic veins.The damage and necrosis of hepatocytes were alleviated in low and medium dose groups,while the structure of hepatocytes was intact in high dose group and positive drug group,and the damage and necrosis of hepatocytes were significantly alleviated.Compared with blank group,the contents of serum TG,TC,AST and ALT in alcohol model group were significantly increased(P < 0.01);Compared with model group,the medium and high dose groups of Penthorum chinense Pursh significantly decreased(P < 0.01)contents of TG,TC,AST and ALT.(3)The different flora of the Penthorum chinense Pursh group was significantly richer than that of the other two groups,and the flora of Codoribacteraceae,Odoribacter,Turicibacter,Turicibacales and Turicibacteraceae were significantly higher than those of the blank group and the model group.In the blank group,Candidatus,Halomonas,Halomonadaceae,Oceanospirillales and Acetobacteraceae were the predominant bacteria groups with high abundance.The bacteria with high abundance in the model group were Clostridia,Clostridiales,Lachnospiraceae,Ruminococcaceae,Oscillospira and Ruminococci Ruminococcus,etc.(4)The important components involved in the treatment of alcoholic liver injury were quercetin,apigenin,kaempferol,luteolin,chonosin,2β,3β,23-trihydroxy-ursolic acid,ursolic acid,ellagic acid,emodin,neocuralin A,etc.Important protein targets include AKT1,EGFR,PIK3R1,MAPK3,PIK3 CA,MET,IGF1 R,RAF1,HRAS,CCND1,etc.The involved disease pathways include cancer,endocrine disorders,prostate cancer,bladder cancer,melanoma,adhesion plaque,non-small cell lung cancer,glioma,pancreatic cancer,etc.The metabolic pathways involved are EGFR tyrosine kinase inhibitor resistance,cancer central carbon metabolism,proteoglycan in cancer,PI3K-Akt signaling pathway,chemocarcinogenic-receptor activation,AGE-RAGE signaling pathway,HIF-1 signaling pathway,Fox O signaling pathway,cancer Micro RNA,Rap1 signaling pathway,Ras signaling Path,etc.Conclusion: 38 components were identified in the 70% ethanol extract of Penthorum chinense Pursh leaves,of which 7 were organic acids and their esters,26 were flavonoids and flavonoid glycosides,and 3 were lignans.The results of pharmacological experiments in mice showed that the alcoholic liver injury model was successfully established,and the symptoms of liver injury were significantly improved in the medium and high dose groups of Penthorum chinense Pursh in a dose-dependent manner.Intestinal flora analysis showed that there were differences in the species diversity and composition of intestinal flora of mice in each group.Penthorum chinense Pursh water extract could regulate the species structure of intestinal flora of mice with alcoholic liver injury,and increase the species and quantity of intestinal beneficial bacteria to improve their richness.Alcohol intake would destroy the balance of intestinal flora of mice.The water extract of Penthorum chinense Pursh can indirectly treat alcoholic liver injury in mice through the "enteric-liver axis".The results of network pharmacologic prediction indicate that the treatment of alcoholic liver injury with Penthorum chinense Pursh is closely related to cancer pathway.