Identification Of Key Pharmacodynamic Substances Of Hedan Capsule In Anti-Nonalcoholic Fatty Liver Disease

Objective:In this study,UPLC-MS,network pharmacology technology,molecular docking technology and zebrafish model were used to preliminarily explore the key drug substances of Hedan capsule in the treatment of nonalcoholic fatty liver,providing scientific basis for the follow-up quality control and clinical application of Hedan capsule.Methods:1 Chemical composition characterization of Hedan capsule(UPLC-QE-MS qualitative analysis).Add 600 ΜL of methanol water(containing L-2-chlorophenylalanine),then grind it in a grinder,ultrasonic extraction in ice-water bath;low-temperature centrifugation,supernatant extraction,pinhole filter filtration,transfer to liquid-phase injection vials,preservation at-80 ° C,until LC-MS analysis.2 Construction of zebrafish nonalcoholic fatty liver disease(NAFLD)model and evaluation of the activity of Hedan capsule in the treatment of zebrafish NAFLD.Select the best concentration and time for the establishment of nonalcoholic fatty liver model of zebrafish,establish the nonalcoholic fatty liver model,and determine the optimal concentration range of Hedan capsule.Within the drug tolerance range,select four dose groups(10,30,50,70)from high to low μ G/ml)to evaluate the activity of Hedan capsule in the treatment of NAFLD.3 Screening of key active ingredients of Hedan capsule in the treatment of nonalcoholic fatty liver.According to 107 chemical components identified by UPLCTOF-MS in the early stage,the target gene of Hedan capsule was collected and the disease target of nonalcoholic fatty liver was obtained from the database.Through multi-level network construction and analysis,the chemical components with the highest degree value of 27(degree>20)were selected as the substances screened for the first time,and then the obtained components passed the content and blood component screening.Subsequently,molecular docking simulation technology was used for further screening,and the components screened in the above steps were used as the active substances preliminarily screened.4 The zebrafish model was used to verify the activity of the active substances preliminarily screened.The zebrafish model was used as the experimental animal to verify the activity of the active substance monomer.Results:1 The database of Hedan capsule was established according to the literature,and107 components and isomers of Hedan capsule were identified by comparison.It mainly includes tanshinones,salvianolic acids,alkaloids,flavonoids,organic acids,phenylpropanoids and anthraquinones.2 According to the experimental results,the conditions for the establishment of nonalcoholic fatty liver model of zebrafish were finally determined as follows: the size of 5dpf AB zebrafish,high-fat diet containing 5% cholesterol,feeding for seven days,and the feed concentration in the system was 0.4mg/ml.Four dose groups(10,30,50,70)were administered with Hedan capsule μ G/ml),50,70 μ The two dosage groups of g/ml showed significant therapeutic effect,which proved that Hedan capsule showed significant therapeutic effect on nonalcoholic fatty liver in zebrafish model.3 According to the results of network pharmacology and molecular docking,the substances that can bind with PPAGR,TP53,AKT1,and IL6 are selected as the active substances of Hedan capsule in the treatment of nonalcoholic fatty liver,namely,lotus leaf alkaloid,psoralen,tanshinone I,tanshinone IIA,ursolic acid,cryptotanshinone,quercetin,and salvianolic acid A.4 According to the zebrafish experiment results,lotus leaf alkaloid,tanshinone IIA,ursolic acid,cryptotanshinone,quercetin and salvianolic acid A showed significant activity in the treatment of nonalcoholic fatty liver.Conclusion: According to the results of this experiment,among the components of Hedan capsule,the key effective substances that play a role in the treatment of nonalcoholic fatty liver are mainly lotus leaf alkaloid,tanshinone IIA,ursolic acid,cryptotanshinone,quercetin,salvianolic acid A.