Network Regulation Mechanism Of Tibetan Medicine DXK Capsule Therapy HAPC Based On Metabolomics

High altitude polycythemia,HAPC,which is called “bad blood plethora” by Tibetan medicine,is a clinical syndrome of a series of symptoms and signs,which caused by erythrocyte compensatory hyperplasia,increased blood viscosity and increased blood flow resistance as the body exposed to plateau hypoxia environment.Duoxuekang capsule(DXK)whose prescriptions was made by Tibetan Master Troru Tsenam on the early clinical basis and indications is blood disease and tripa disease has curative effect of clinical treatment to Plethora.At present,due to the lack of research on the mechanism of molecular metabolism level,in the treatment of HAPC with DXK capsule,we can not solve what is the difference in components between normal blood and "bad blood" and DXK capsule how to change the composition of the "bad blood".In this study,we mainly used the methods of network pharmacology and metabonomics to study the mechanism of DXK capsule in the treatment of the HAPC.Purpose:1.Investigating the effect of the treatment of DXK capsule to HAPC in rats;2.Investigating the mechanism of HAPC in the treatment of DXK capsule.Methods:1.Using network pharmacology method,screening compounds from the chemical composition of DXK capsule by HPLC research group in the early;applying PharmMapper server for potential target prediction,targeting specification name;the potential target is imported to MAS 3.0 database with KEGG database for circuit analysis,finally using Cytoscape software to construct DXK capsule "active ingredient-target-path” network diagram.2.Pharmacodynamic study on the treatment of HAPC model rats with the DXK capsule: use low pressure chamber model to establish the HAPC model rats,with intragastric administration of different doses of the capsule.Through the detection of blood routine,hemorheology,LDH,SDH,EPO and other indicators of model rats,combined with pathological observation,we observe the therapeutic effect of HAPC in the treatment of the capsule.3.Metabonomics study on the treatment of HAPC with DXK capsule:in the serum of HAPC model rats as the research object,basing on UHPLC-Q/TOF-MS analysis technology and combining of multivariate statistical analysis techniques to analyze and process data,we investigate the potential biomarkers and differential metabolites of disease.Integrate and analyze the potential differences in biomarker of the different groups,construct the metabolic pathway network and analyze the potential mechanism of the treatment of HAPC with DXK capsule.Results:1 After network pharmacology analysis,42 chemical constituents,54 targets and56 pathways were found,It mainly targets CDK2,AKT1,GSK3β and its most significant pathway are Adipocytokine signaling pathway,Complement and coagulation cascades,Glycine serine,and threonine metabolism,Tryptophan metabolism,VEGF signaling pathway.2.The hematological indexes of HAPC model rats showed that RBC,HGB,HCT,whole blood hemorheology were all increased in different degree,but the hematology indexes in the treatment group were decreased to some extent.The detection of hypoxia related cytokines showed that DXK capsule could improve the activity of SDH and decrease the expression of the activity of LDH、EPO.The results of pathological section showed that DXK capsule can significantly reduce the damage of HAPC model rats’ lung and heart tissue by altitude hypoxia.It is suggested that DXK capsule has a good protective effect on tissue damage caused by hypobaric hypoxia.3.Metabonomics experiments has obtained 37 metabolic difference in serum of HAPC model rats,Hydroxyproline,Oxidized glutathione,Pantothenate etc of the metabolic difference have a significant difference,involving metabolic pathways including D-glutamine and D-glutamate metabolism,Alanine,aspartate and glutamate metabolism.After DXK capsule treatment,16 characteristic differential metabolites are obtained,including Oxidized glutathione,Succinic acid,Glyceryl acid etc,involving Ketone synthesis and degradation,Glycerolipid metabolism,etc.Thereare 4 metabolic pathway soverlaping with HAPC pathogenesis related metabolic pathway.Conclusion:1 DXK capsule can reduce the number of red blood cells,reduce blood viscosity.At the same time,the capsule has obvious anti hypoxia effect,can improve the heart,lung due to pathological changes caused by hypoxia,show DXK capsule is effective.2 The pathogenesis of HAPC may be related to energy metabolism,oxidative stress metabolism,lipid metabolism,intestinal metabolic metabolism and other related metabolites and pathways.DXK capsules treat HAPC by affecting the immune response,inflammatory response,oxidative stress and other targets of the body and correcting the above-mentioned metabolites and metabolic pathways.By using the network pharmacology and metabolomics technique,revealing the mechanism of "bad blood" in Tibetan medicine theory,and the molecular mechanism of DXK capsule to correct "bad blood",to provide scientific support for the clinical treatment of the disease mechanism of Tibetan Medicine.