Study On The Material Basis Of Antidepressant Effect Of Bupleurum Scorzonerifolium And Vinegar Bupleurum Scorzonerifolium

Bupleurum scorzonerifolium is the dried root of Umbelliferae plant Bupleurum scorzonerifolium Willd..and it is one of the sources of Bupleurum chinense collected in Pharmacopoeia of the People’s Republic of China(2020 edition).It is mainly produced in Heilongjiang,Jilin,Hubei and other provinces,with the efficiency of relieving fever,soothing the liver and relieving depression,lifting Yang Qi.According to the literature,the antidepressant effect of B.scorzonerifolium is better than that of Bupleurum chinense.Vinegar slices of Bupleurum chinense are usually used in the clinical treatment of depression in Chinese medicine.However the material basis of antidepressant effect of B.scorzonerifolium and the changes of chemical components before and after processing are still unclear.In this study,a rat model of depression was established to evaluate the difference of antidepressant effect of B.scorzonerifolium before and after vinegar processing.Then chemical constituents and blood entry components of B.scorzonerifolium and Vinegar B.scorzonerifolium,were further analyzed.Network pharmacology was applied to explore the material basis of antidepressant effect and to screen antidepressant quality markers.The results will lay a foundation for the quality evaluation of B.scorzonerifolium medicinal materials,prepared slices and related preparations.First,the antidepressant pharmacodynamics of B.scorzonerifolium and vinegar B.scorzonerifolium were studied.The depression model was established by CUMS combined with orphaning method.After 28 days,the body weight,sugar preference,open field activity score and the contents of 5-HT,DA and NE in the brain tissue of the model rats were lower than those of rats in the normal control group(P<0.05),indicating that the depression model was established successfully.After,B.scorzonerifolium,vinegar B.scorzonerifolium and positive drug were given to depressed rats for 21 days,the body weight,sugar preference,open field activity score and the contents of 5-HT,DA and NE in the brain tissue of rats were much higher than those of rats in the negative control group(P<0.05).The antidepressant effects of vinegar B.scorzonerifolium was similar to B.scorzonerifolium.Although vinegar B.scorzonerifolium all index of vinegar B.scorzonerifolium was slightly better than that of B.scorzonerifolium there was no statistic difference(P>0.05).Secondly,The chemical constituents of B.scorzonerifolium and vinegar B.scorzonerifolium were analyzed,UPLC-Q-TOF-MS/MS was used for qualitative analysis.combined with the self-bult chemical constituent database and reference substance database.A total of 39 compounds were identified,in which 38 compounds were found in B.scorzonerifolium and vinegar B.scorzonerifolium and ProsaikogeninA/D/G/F were only found in vinegar B.scorzonerifolium.13 compounds with different contents were found.The contents of SSa,SSc,SSd,Isorhamnetin-3 murine O-β-D-glucoside,Isochlorogenic acid,SSk,SSp,Isorhamnetin and Daffodil decreased after processing.The contents of SSb1,SSb2,Fraxetin and Petunidin increased after processing.Third,the blood components of B.scorzonerifolium.and vinegar B.scorzonerifolium.were analyzed in depressed rats.UPLC-Q-TOFMS/MS was used to analyzed components in the blood of depressed rats.4 prototype components(Valerenic Acid,Nootkatone,DIBP,Glycyrrhetinic acid)and 2 metabolites(Saikogenin A/D/F/G,Saikogenin E)were indentified.There was no difference in blood components between B.scorzonerifolium.and vinegar B.scorzonerifolium..Saikogenin A,D,G,F are isomers.Saikogenin A is the metabolite of SSa or SSb1.Saikogenin D is the metabolite of SSd or SSb2.Saikogenin F is the metabolite of SSa.Saikogenin G is the metabolite of SSd.Saikogenin E is the metabolite of SSc.Therefore,9 compounds(SSa,SSb1,SSb2,SSc,SSd,Valerenic Acid,Nootkatone,DIBP and Glycyrrhetinic acid)were proposed as the potential antidepressant material basis of B.scorzonerifolium.and vinegar B.scorzonerifolium..Fourth,9 potential antidepressant substances were analyzed by Network Pharmacology.The targets of SSb1,SSb2,SSd,SSa,SSc,ValerenicAcid,Nootkatone,DIBP and glycyrrhetinic acid were collected from SwissTargetPrediction database and Pharmmapper database,and the composition-target network diagram was constructed.Then using the String website to analyze the interaction between targets,16 core targets are obtained,and the target interaction network diagram is constructed.Finally,the core targets were enriched and analyzed by DAVID database,and 20 signal pathways were obtained.Combined with the literature,PI3K-Akt,Ras and MAPK signaling pathways were closely related to antidepressant mechanism.SSa,SSb1,SSb2,SSc,SSd,Nootkatone and Glycyrrhetinic acid were further confirmed as antidepressant substances of B.scorzonerifolium.and vinegar B.scorzonerifolium..Finally,the contents of SSa,SSc and SSd were selscted as the antidepressant quality markers in 7 antidepressant active compounds,because of their stability,biological activity,high content and easily obtaining reference substance.The content of SSa,SSc and SSd in B.scorzonerifolium.and vinegar B.scorzonerifolium.from 6 different origins were determined by HPLC-ELSD.The total contents of SSa,SSc and SSd in 6 batches of B.scorzonerifolium.from different origins ranged from 0.0734%to 0.1414%.The total contents of SSa,SSc and SSd in 6 batches of vinegar B.scorzonerifolium.from different origins ranged from 0.0391%to 0.0764%.To sum up,the antidepressant substances of B.scorzonerifolium.and vinegar B.scorzonerifolium.are SSa,SSb1,SSb2,SSc,SSd,Nootkatone and Glycyrrhetinic acid.It is proposed that SSa,SSc and SSd could be used as antidepressant quality markers of B.scorzonerifolium.and vinegar B.scorzonerifolium..The results provide a scientific basis for the quality standard establishment of B.scorzonerifolium.and vinegar B.scorzonerifolium.prepared slices and related preparations.