Based On Network Pharmacology To Analyze The Prevention Mechanism Of Hugan Tablet On Alcoholic Liver Injury

Objective: Since 1980,my country’s wine culture has gradually become popular,and alcohol consumption has increased rapidly.Excessive drinking has become an important factor causing a series of social and economic problems,and the alcoholics have gradually become younger.Alcoholic liver injury(ALD)seriously harms human health and brings serious medical and economic problems to society.Acute alcoholism and long-term alcoholism can cause damage to various organs and tissues,especially liver damage.In view of the above situation,it is pressing to seek drugs to guard against and cure alcoholic liver injury.In order to find drugs,we focused on traditional Chinese medicine.HGP contains six ingredients,mung bean of proprietary Chinese medicine,with liver qi,spleen to fall the action of enzymes and cholagogue lipid-lowering,widely used in clinical chronic hepatitis,control and prevention of early cirrhosis clinical also widely used in the treatment of ALD,but its mechanism of action of ALD is unclear.HGP has the effects of dredging liver,regulating qi,strengthening spleen and lowering enzyme,enhancing bile and lowering lipid.It is widely used in the prevention and treatment of chronic hepatitis,drug-induced liver injury and early cirrhosis,and also used in the treatment of ALD clinically,but its prevention and treatment mechanism of ALD is still unknown.Traditional Chinese medicine refers to the Chinese medical way,in the Spring and Autumn Period and the Warring States Period,it formed foundation,through the development of dynasties,it formed shaping as today.Modern study shows that a single Chinese herbal medicine has several components and targets,and the prescription composed of multiple Chinese herbs plays a role by the mechanism of multiple pathways corresponding to its multiple targets.Application of network pharmacology in traditional Chinese medicine: through mining the effective active ingredients and target prediction of traditional Chinese medicine,network diagram analysis of their relationship,from the molecular level to elaborate the possible mechanism of the combination of traditional Chinese medicine prescription to exert efficacy.In this paper,Hugan Tablet was taken as the research object to analyze its mechanism of action in the treatment of alcoholic liver injury and to verify it.Methods: 1.Based on the results of network pharmacological prediction,using Western Blot and Real-time PCR to validate result,and the contents of relevant inflammatory factors in serum were determined.The protein and m RNA levels of NLRP3,caspase-1,interleukin-1β and interleukin-18 in liver tissue of each group were detected.Using Elisa to observe the changes of IL-18 and IL-1β in serum.2.The HGP of ALD in mice liver protection function assessment: acute alcoholic liver injury in mice Model was established and subacute alcoholic liver injury Model in mice,divided into the Control group,Model group and HGP,groups of experimental animal serum AST(Aspartate aminotransferase),ALT(Alanine aminotransferase),ALP(alkaline phosphatase),TP(Total Protein),propagated,TG(Triglyceride),TC(Total cholesterol),LDC-L(Low-Density Lipoprotein Cholesterol),HDC-L(High-Density Lipoprotein)value and anti-oxidation indexes of MDA(Malondialdehyde),GSH(Glutathione)and SOD(Superoxide dismutase)value,at the same time for the liver tissue in HE staining experiment,observed the pathological changes of mice liver tissue.3.Network pharmacology analysis that HGP can be used of alcoholic liver injury:First,use the TCMSP to screen the active components and corresponding targets of HGP;use Cytoscape 3.2.1 software to construct a network relationship diagram between components and targets;The targets were analyzed by GO through Metascape and KEGG pathway by David online database to predict the potential mechanism of HGP in the treatment of alcoholic liver injury.Results: 1.Analysed the potential pathways of HGP in the treatment of alcoholic liver injury based on network pharmacology: a total of 147 active ingredients were screened out from six Chinese herbal medicines,corresponding to 351 targets.There were 249 biological processes involved in drug metabolism,amino acid metabolism,antioxidant process and nervous system related process and other four categories.KEGG was enriched into 52 signaling pathways,involving amino acid metabolism related pathways,immune inflammation related pathways,antioxidant related pathways,nervous system related pathways,cancer and 6 other categories.Among them,inflammation and amino acid metabolism pathways are the main part.The NOD pathway with the highest score was involved in the underlying inflammatory pathway.2.HGP had the effect of treating acute alcoholic liver injury: HGP could significantly reduce the contents of AST,ALP and ALT in serum liver function of Model group(*P< 0.05 or **P < 0.01);The lipid metabolic disorders were improved in HGP group,and the indexes of total cholesterol(TC),triglyceride(TG)and low density lipoprotein(LDL-C)were significantly decreased(**P < 0.01 or *P < 0.05).The contents of antioxidant indexes glutathione(GSH)and superoxide dismutase(SOD)were significantly increased(**P < 0.01),and the content of peroxide malondialdehyde(MDA)was significantly decreased(**P < 0.01).He staining showed that liver fat lesion was improved obviously,lipid droplets in liver cells decreased,and cytoplasm tended to the normal group.3.HGP has the effect of treating subacute alcoholic liver injury: HGP can significantly reduce the serum liver function ALP,AST and ALT(*P < 0.05),and significantly increase the contents of TP and albumin ALB in subacute Model group(*P < 0.05 or **P < 0.01).The contents of TC and LDL-C,the lipid metabolic indexes,were significantly decreased(**P < 0.01 or *P < 0.05),and the contents of TG and HDL-C were significantly increased(**P < 0.01 or *P < 0.05).The contents of GSH and SOD in HGP group were significantly increased(**P < 0.01 or *P <0.05),and the content of MDA was significantly decreased(**P < 0.01).Compared with the Model group,the liver cells in the HGP group had no obvious fat droplets,no obvious infiltration of inflammatory cells,and the liver cells were arranged neatly,and the liver damage state was significantly improved.4.HGP can reduce the inflammatory response of alcoholic liver injury: HGP can significantly reduce the content of NLRP3 inflammasome in acute and subacute Model groups(**P < 0.01),and then decrease the contents of Caspase-1,IL-1β and IL-18(**P < 0.01 or *P < 0.05).Conclusion: 1.HGP can significantly improve the elevated transaminase,lipid metabolism disorder,hepatic fatty disease and inflammation caused by acute and subacute alcoholic liver injury.2.Based on network pharmacology,it was analyzed and predicted that the treatment of alcoholic liver injury by HGP was related to the reduction of the expression levels of NLRP3,Caspase-1,IL-1β and IL-18 in NOD pathway.