Study On The Chemical Material Basis Of Xueshuan Xinmaining Tablets For Blood-Activating And Stasis-Resolving

Based on the research progress of Xueshuan Xinmaining Tablets(XXT)and the review of research methods of the chemical material basis of traditional Chinese medicine,the chemical material basis of XXT for blood-activating and stasisresolving was comprehensively studied by using some methods including chemical composition analysis,spectrum-effect relationship,serum pharmacochemistry and network pharmacology.The innovative results are as follows:1.Analysis and identification of chemical components of XXT based on UNIFI natural product analysis platformUltra performance liquid chromatography quadrupole-time of flight mass spectrometry(UPLC-Q-TOF/MS)technology combined with the UNIFI natural product analysis platform were used for the first time to quickly analyze and identify the chemical components(with the molecular weights from 100 to 1500 Da)from70% methanol extract of XXT.A total of 187 chemical constituents,including triterpenoid saponins,phenanthrene-quinones and steroids and so on,had been identified by comparing with reference substances,or by analyzing accurate molecular weight typical fragments.It was demonstrated that XXT was rich in the chemical components with various structural types.The structural diversity of chemical components in XXT was the chemical material basis for the multi-target pharmacological effects.2.Screening directional efficacy components of XXT for blood-activating and stasis-resolving based on spectrum-effect relationshipFirstly,the HPLC fingerprints of XXT different polarities solvent extracts were established for the first time,and the activities of each extract for blood-activating and stasis-resolving were evaluated.Then,the spectrum-effect relationship between peak areas in chromatograms and the indexes of anti-blood stasis was established bymultivariate statistical analysis,including gray relational analysis and partial least squares regression analysis.A total of 9 candidate components in XXT were screened out.Among them,7 components including salvianic acid A(peak 1),rutin(peak 3),ginsenoside Rg1(peak 11),ginsenoside Rb1(peak 22),cinobufagin(peak 36),tanshinone I(peak 38),and tanshinone IIA(peak 39)were verified to be the potential directional efficacy components of XXT for blood-activating and stasis-resolving by the validation experiment in vitro.3.Identification of migrating components of XXT in serum and brain tissue of blood stasis rats based on serum pharmacochemistryBased on UPLC-Q-TOF/MS technology combined with multivariate statistical analysis,including principle component analysis and orthogonal partial least squares discrimination analysis,the study on components of XXT migrating to serum and brain tissue of blood stasis rats was performed for the first time.(1)Identification of migrating components of XXT in serumA total of 30 prototype constituents,such as triterpenoid saponins,phenanthrenequinones and steroids,had been identified for the first time by comparing with reference substances,or by analyzing accurate molecular weight and typical fragments.Among them,the 14 and 28 components were identified from healthy rats and blood stasis rats,respectively.Only 12 shared components were found.Since the contents of ginsenoside Rd,cryptotanshinone,dihydrotanshinone I,cholic acid,taurodeoxy-cholic acid,Ilexgenin A,and Senkyunolide H were higher and only were detected in serum of blood stasis rats.It was inferred that these 7 ingredients were the biologically active ingredients of XXT in blood stasis rats.(2)Identification of migrating components of XXT in brain tissueA total of 11 prototype constituents had been identified for the first time by comparing with reference substances,or by analyzing accurate molecular weight and typical fragments.Among them,the contents of 19-Hydroxybufalin and cinobufagin-3-suberoyl ester were higher than other components.Therefore,it was inferred that the 2 ingredients were the biologically active ingredients of XXT in blood stasis4.Establishing networks of "XXT chemical material basis-blood stasis targets-pathways" based on network pharmacologyA total of 16 chemicals of directional efficacy components and migrating components of XXT were chosen as "candidate compounds".The interaction network of "XXT chemical material basis-blood stasis target-pathway" was constructed by network pharmacology technology for the first time.The potential key targets(STAT3,VEGFA,AKT1,TNF,IL6,and MMP9,etc.)and the related signaling pathways(proteoglycans in cancer,PI3K-Akt,TNF,Rap1 and NF-?B,etc.)were predicted.The intervention and impacts of 16 chemical materials on the disease network were explored from a holistic perspective.The results also theoretically verified the synergistic mechanism of all the chemicals screened out by the spectrum-effect relationship analysis in vitro and the serum pharmacochemistry in vivo.In summary,a total of 16 chemicals of XXT for blood-activating and stasis-resolving were identified based on studies in vivo and in vitro.This dissertation would provide detailed scientific data for elucidating the chemical material basis of this traditional Chinese medicine,and also provide new idea for the chemical materials study of traditional Chinese medicine.