Based On The Systemic Pharmacology Method, The Mechanism Of Action Of Traditional Chinese Medicine Yiqi Capsule In Treating ITP Was Studied

Primary immune thrombocytopenia(ITP),also known as idiopathic thrombocytopenic purpura,is caused by thrombocytopenia which results from the loss of immune tolerance to the patient’s own platelet antigens.In patients with ITP,megakaryocyte maturation disorders occur in bone marrow.And ITP patients may have the serious consequence with or without bleeding into the skin,mucous membranes and internal organs and even can be life-threatening.At present,Western medicine is limited and has many side effects in ITP treatment.Therefore,there is an urgent need to speed up the research process that traditional Chinese medicines treat ITP.Yiqi Shexue formula is composed of Panax ginseng C.A.Mey.,Poria cocos(Schw.)Wolf,Atractylodes macrocephala Koidz.,Glycyrrhiza uralensis Fisch,Angelica sinensis(Oliv.)Diels,Ligusticum chuanxiong Hort.,Paeonia lactiflora Pall.,Equus asinus L.,and Rehmanniae Radix Praeparata.Yiqi Shexue formula is a plus-minus formula of Bazhen decoction and has effects on invigorating spleen,replenishing qi,nourishing the blood and promoting blood circulation.It is mainly used to treat ITP.Bazhen decoction is a representative formula of tonifying qi-blood.Pharmacological investigations have shown that Bazhen decoction has the effects of improving hematopoietic function,anti-platelet aggregation,improving hemorheology,and improving the body’s immunity.However,the mechanisms and material basis of Yiqi Shexue formula is not clear and the related research is lacking.The objective of this study is investigating the material basis and mechanisms that Yiqi Shexue formula treats ITP.Firstly,the characteristic chromatogram of Yiqi Shexue formula was constructed using an ultra performance liquid chromatograph in the study.Then constituents absorbed into blood and spleen was investigated through LC-MS/MS technology.Finally,the mechanisms that Yiqi Shexue formula treats ITP were explored through a network pharmacology approach.Results of the study are conducive to revealing the mechanisms that Yiqi Shexue formula treats ITP and provide a new way for the research of material basis and mechanisms of traditional Chinese medicine.Methods:1.Study in chemical components of Yiqi Shexue formula:Yiqi Shexue formula was dissolved in methanol and the sample was prepared after ultrasonic treatment.The chemical components of Yiqi Shexue formula were authenticated by comparing relative retention time and maximum absorption wavelength of reference substances using UPLC.2.Study in constituents absorbed into blood and spleen:Yiqi Shexue formula was infused to stomach of SD rats in a high dose(4.14g/kg)twice daily for 3 days.The plasma and spleen of rats were detected by LC-MS/MS technology after a series of preliminary treatment.The constituents absorbed into blood and spleen of Yiqi Shexue formula were authenticated by comparing retention time.3.Study in the mechanisms that Yiqi Shexue formula treats ITP:The constituents of Yiqi Shexue formula absorbed into blood and targets of ITP were studying.And the compounds-targets network and protein-protein interaction network of Yiqi Shexue formula,protein-protein interaction network of ITP-related targets,and targets network that Yiqi Shexue formula treats ITP were constructed by a network pharmacology approach.The mechanisms that Yiqi Shexue formula treats ITP were explored by carrying out GO and KEGG analyses.Results:1.The characteristic chromatogram of Yiqi Shexue formula was constructed by UPLC analysis and 10 compounds were authenticated,namely:gallic acid,oxypaeoniflora,chlorogenic acid,albiflorin,paeoniflorin,ferulic acid,verbascoside,senkyunolide I,senkyunolide H and benzoylpaeoniflorin.2.Twenty-three compounds of Yiqi Shexue formula were identified to be absorbed into blood and 11 compounds were identified to be absorbed into spleen by UPLC-MS/MS analyses.23 compounds absorbed into blood are ginsenoside Rbl,ginsenoside Re.ginsenoside Rd,ginsenoside Rc,ginsenoside Rb2,ginsenoside Rb3,ginsenoside Rgl,rehmannioside D,catalpol,5-hydroxymethyl-2-furancarboxylic acid,ajugol,isoliquiritigenin,isoliquiritin,glycyrrhetnic acid,pachymic acid,paeoniflorin,oxypaeoniflora,senkyunolide Ⅰ,senkyunolide H,ferulic acid,chlorogenic acid,3-butyl-1(3H)-Isobenzofuranone,and atractylenolide Ⅲ.11 compounds absorbed into spleen are ginsenoside Rbl,ginsenoside Re,ginsenoside Rd,ginsenoside Rc,ginsenoside Rb2,ginsenoside Rb3,ginsenoside Rgl,isoliquiritin,senkyunolide I,and ferulic acid.3.The active compounds of Yiqi Shexue formula and theirs targets were analyzed using a network pharmacology approach and the mechanisms that Yiqi Shexue formula treats ITP were explored by carrying out GO and KEGG analyses.Some biological process including negative regulation of gene expression,cellular protein metabolic process,and cell adhesion were enriched.Cellular component such as nucleoplasm,cytoplasm,and protein complex and molecular function including protein binding,ubiquitin protein ligase binding,and transcription factor binding were enriched.In KEGG analyses,signaling pathways such as neurotrophin signaling pathway,B cell receptor signaling pathway,PI3K-Akt signaling pathway,MAPK signaling pathway and NF-kappa B signaling pathway were enriched.Conclusion:The characteristic chromatogram of Yiqi Shexue formula was constructed in the study,which provides a method to control the quality of Yiqi Shexue formula.Twenty-three compounds of Yiqi Shexue formula were identified to be absorbed into blood and 11 compounds were identified to be absorbed into spleen,which can lay the foundation for further research about the pharmacodynamic material basis of Yiqi Shexue formula.Results of network pharmacology research indicated that Yiqi Shexue formula might have a role in treating ITP by regulating some biological processes including gene expression,cellular protein metabolic process,and cell adhesion,and involving some cellular component,such as nucleoplasm,cytoplasm,and protein complex.The mechanisms might be concerned with some signaling pathways,such as neurotrophin signaling pathway,B cell receptor signaling pathway,and PI3K-Akt signaling pathway.