Discovery And Research On Active Ingredients Of Traditional Chinese Medicine For Prevention And Treatment Of Liver Injury

The liver is not only an important organ responsible for the metabolism of energy and substances in the body,but also an important detoxification organ.The incidence of liver diseases caused by various hepatotoxic substances has increased year by year.Alcohol and drugs are the most common liver poisons.Beasuse more than 90%of alcohol is oxidized and metabolized in the liver,the liver is the most serious organ affected by alcohol injury.Druginduced liver injury is the most common clinical adverse drug reaction.Therefore,the discovery strategy of hepatoprotective drugs from traditional Chinese medicine for the treatment of alcohol-induced and drug-induced liver injury has become the focus of drug development.It was found that Alcohol dehydrogenase(ADH)is an important therapeutic target for alcohol-induced liver injury,and Farnesoid X receptor(FXR),Liver X receptor(LXR?),pregnane Pregnane X receptor(PXR),Peroxisome proliferators-activated receptor ?(PPAR?),Proteinase-activated receptor 1(PAR-1)are important therapeutic targets for drug-induced liver injury.In this research,the combination of molecular simulation and biological experiments was utilized to explore the hepatoprotective compounds from traditional Chinese medicine.What is more,an exploratory research about 'Target-Acitivity Relationship' was proposed for the overall efficacy evaluation of drugs based on multi-target.Firstly,the interacion between the potential hepatoprotective compounds and targets were obtained by molecular simulation method.Then,the hepatoprotective activity of the potential hepatoprotective compounds were achieved by biological experiment.The target-acitivity relationship was studied machine learning method,which provides guidance for the overall efficacy evaluation of compounds based on multi-target.1.Researchment on Hepatoprotective TCM Compounds for Alcohol-induced Liver Injury Based on Alcohol Dehydrogenase InhibitorsIn this research,pharmacophore and molecular docking screening models for ADH inhibitors were constructed to screen the potential ADH inhibitors.Then,the inhibitory activities of these compounds were screened by the ADH enzyme activity assay.L33,an ADH inhibitor was obtained with the IC50 of 1.926 ?mol·L-1.Based on the Lineweaver-Burk plot,L33 displayed a competitive inhibition pattern.The interaction mode between L33 and ADH1 family A,B,C three subtype proteins was analyzed by molecular dynamic method.Finally,the hepatoprotective activity of L33 against the alcohol-induced liver injury was experimented on L02 cells.The study found that 12.5-50 ?mol·L-1 L33 had a significant protective effect on alcohol-induced liver injury.2.Researchment on Hepatoprotective TCM Compounds for Drug-induced Liver InjuryBased on the molecular docking models of FXR,LXR-?,PAR-1,PPAR-?,and PXR,112 potential hepatoprotective TCM compounds were obtained.Then,the effects of 112 potential hepatoprotective TCM compounds on the normal L02 cells at 50 ?mol·L-1 were detected.Only 58 potential compounds with L02 cell survival rate higher than 90%were retained for the test of its hepatoprotective activity by the Acetaminophen(APAP)-induced liver injury cell model.It was found that M17,M43,M55 protect against acetaminophen-induced hepatotoxicity at 50?mol·L-1.3.Researchment on Mechanism of Hepatoprotective Compounds for Drug-induced Liver InjuryBy examining the effects of the compounds on the normal L02 cells and APAP-induced liver injury L02 cells,it was found that M17,M43,and M55 have significant protective effects on APAP-induced liver injury L02 cells.M17,M43,and M55 have an inhibitory effect on the increase of AST,AKP,and ROS in APAP-induced L02 cell injury.M17,M43 can promote the decrease of GSH in L02 cells.By detecting the gene expression of targets related to druginduced liver injury,it was found that M17,M43,M55 can increase the expression of FXR,PPAR-?,LXR-? gene in APAP-induced liver injury cells,respectively.The above results indicate that M17,M43,and M55 have therapeutic effects on APAP-induced liver injury,and can regulate the abnormality of AST,AKP,GSH,and ROS caused by APAP.Their therapeutic effects may be related to the activation of FXR,PPAR-? and LXR.4.Researchment on Target-acitivity Relationship of Drug-induced Liver InjuryThe interaction between the 58 potential compounds and 5 targets related to drug-induced liver injury were defined as independent variable.The hepatoprotective activity of the 58 potential compounds was defined as dependent variable.MLP algorithm,SVR algorithm,DTR algorithm,GBR algorithm were utilized for the research of target-acitivity relationship of druginduced liver injury.The effect of different activation algorithm and different slover for MLP models were compared.SVR models were constructed by different kernel functions.Different feature selection criterion and splitter for DTR models and different number of estimators and loss functions for GBR models were also considered in this research.According to the results,GBR target-activity relationship model achieved high fitting accuracy and good prediction ability.Then the optimal GBR target-activity relationship model was utilized to screen the traditional Chinese medicine database for the potential hepatoprotective TCM compounds.In this study,L33 was discovered as an inhibitor of ADH and can reduces the alcohol-induced liver injury.The TCM compounds M17,M43,and M55 were discovered as the hepatoprotective activity against drug-induced liver injury by activating of FXR,PPAR-?,and LXR.Moreover,an exploratory research about 'Target-Acitivity Relationship' was proposed for the overall efficacy evaluation of drugs based on multi-target.The relationship between the compounds'interaction with targets and hepatoprotective activity were explored in this research.It may provide reference for the overall efficacy evaluation of compounds and development of hepatoprotective TCM drugs.