Synchronous Study On The Effective And Toxic Substances In Tripterygium Glycosides Tablets

Objective: This research simultaneously focused on the effective and toxic ingredients in Tripterygium glycosides tablets in the treatment of rheumatoid arthritis.Methods: The anti-rheumatoid arthritis and hepatotoxic substances in Tripterygium glycosides tablets were studied by the integration of metabolomics,network pharmacology,and in vivo chemical composition analysis.In present study,the endo-and exogenous substances in rat serum were analyzed at different time points after administration of Tripterygium glycosides tablets.1)The changes in serum metabolites at different time points were dynamically processed by non-target metabolomics,then TGT targets were predicted using metabolomics combined with network analysis.RA targets and hepatotoxicity targets were obtained from online database.The potential effective and toxic targets of TGT were obtained from the intersections of TGT targets,RA targets and hepatotoxicity targets.2)The TGT ingredients in rats were identified by UHPLC-Q-TOF-MS technology.Three networks of compounds-effective/toxic targets were constructed using the compounds in TGT in vitro or in vivo,RA and hepatotoxicity targets and potential effective and toxic targets of TGT.The effective and toxic substances in TGT were obtained simultaneously.Animal models with different pathological stages were established by treating CIA rats at different doses of TGT.The efficacy of TGT was evaluated by sole diameter,arthritis score,CT and so on.The hepatotoxicity of TGT was evaluated by liver coefficient and morphology,histopathological staining and so on.Metabolic profile and differential metabolites were analyzed by non-target serum and liver metabolomics.To screen out potential effective or toxic biomarkers,statistical analysis was performed to model associations of differential metabolites with effective and toxic index.The TGT ingredients in CIA rats were identified by UHPLC-Q-TOF-MS technology.Then,the effective and toxic substances in TGT were screened out by correlation analysis between effective/toxic index/biomarkers and in vivo substances in TGT.Results: In normal rats,3 targets of the treatment of RA were found,including SNRNP70,SNRPD3,CRLF1,SGPL1,RELN,PLA2G1 B,PTGES,PLA2G10.5 targets of hepatotoxicity were obtained,such as DCTN1,FASN,LRAT,PRKCA,ASAH1,ACSL1,PLA2G6,PLA2G7,RBP1,TUBA1 A,APOC3.APOA1,APOB,LCAT,ALOX5,PTGS2,PTGS1,PLA2G4 A,PLA2G2A,STAT3 were the targets of both therapeutic and hepatotoxicity.A total of 33 constituents in Tripterygium glycosides tablets were identified in normal rats.Combined with network pharmacology,5 alkaloids had hepatotoxicity,including 1-Desacetylwilforgine,Wilfordconine,Wilforgine,Wilformine,Wilfornine D;and 8 ingredients were found to be effective and toxic,including 14-oxo-19-(4→3)abeo-abieta-3,8,12-tetraen-19,18-olide,7-oxo-18(4→3)abeo-abieta-3,8,11,13-tetraen-18-oic acid,Hypoglaulide,Triptotriterpenic acid A,Wilforol F,Wilforlide B,Triptoquinone B,Wilforlide A.In CIA rats,14 potential therapeutic biomarkers and 25 potential hepatotoxic biomarkers were screened by the integration of metabolomics and statistics.A total of 96 components were found in the serum of CIA rats,23 of which were successfully identified.According to the results of correlation analysis,the therapeutic effect and hepatotoxicity of 54 ingredients in vivo were evaluated.The results shown that 2 compounds shown therapeutic effects and non-toxic,7 components present hepatotoxicity but non-therapeutic effect,42 components have both therapeutic effect and hepatotoxicity,3 compounds shown non-toxic and non-therapeutic effects.Conclusion: This study integrated metabolomics,network pharmacology and chemical compounds analysis to simultaneously discovery and screen the effective and toxic substances of Tripterygium glycosides tablets.Combined with the two results,2 ingredients had therapeutic effect and non-toxic,12 substances were only shown hepatotoxicity,48 components presented both therapeutic effect and hepatotoxicity,3 components presented no therapeutic effect or hepatotoxicity.