| Metabolic syndrome(MetS)is a complex pathological condition in which multiple cardiovascular risk factors such as central obesity,insulin resistance,dyslipidemia,and hypertension are concentrated.The main clinical consequences of MetS are cardiovascular and cerebrovascular diseases and type 2 diabetes,which have been globally identified as a major health issue that affects human health.At present,insulin resistance is the common pathophysiological basis of MetS.Central obesity is an important factor causing insulin resistance,but its exact pathogenesis still remains vague.There is still no specific treatment for MetS.Traditional Chinese herbs have many merits such as organic chemical components,multiple action pathways and synergistic effects of multiple targets.It has its own characteristics and advantages in the prevention and treatment of complex diseases,and is expected to become an important strategy for the prevention and treatment of MetS.The classic famous formula named Wendan decoction(WDD),which was clinically applied in multi-system complex diseases such as neurological diseases,digestive diseases,hypertension,diabetes,obesity and other MetS-related diseases,and its clinical curative effect was remarkable.However,the current organic components and action mechanism of WDD against MetS still remain unclear,and the overall pharmacological action characteristics of the multi-organic chemical component-multi-target-multi-pathway in vivo also have not been revealed.Considering the fact that there is still no research method to fully understand the pharmacodynamic substances and action mechanism of traditional Chinese herbs,this thesis comprehensively adopted multidisciplinary methods such as chemical informatics and metabolomics to reveal the pharmacological organic components and action mechanism of WDD against MetS from levels of chemical information,animals,cells,proteins and metabonomics,which provided evidence for clinical drug selection and efficacy evaluation and also new ideology and methods for understanding the organic components and mechanism of other Chinese herbs.The main research work was as follows:First,we applied network pharmacology methods such as chemical informatics,principal component analysis,molecular docking and network analysis to perform data collection,database construction,drug-like analysis,molecular target identification and network construction of organic components-molecular targes-biological pathways in order to screen active organic compounds,target proteins and biological pathways against MetS from hundreds of organic chemical constituents of WDD and provide guidance for WDD chemical substances and pharmacodynamic experimental research.As a result,it was revealed that 217 active organic compounds of WDD had strong binding affinity to PPAR?,PPAR?,PPAR?,LXR?,LXR? and RXR?,which were invovled in 19 biological pathways.The main pharmacological effects of WDD were involved in maintaining lipid and glucose metabolism balance,anti-cytotoxicity,immune regulation and liver protection.Second,we applied UHPLC-Q Exactive Orbitrap/MS technology to rapidly separate and qualitatively identify 110 organic compounds targeting metabolic nuclear receptors in WDD with good water solubility and intestinal absorption.The chemical structures of 36 organic compounds were inferred based on UHPLC-Q Exactive Orbitrap/MS detection of primary/secondary mass spectrometry data and self-built WDD organic compound chemical information database,and also combined with ACD Lab/MS Fragmenter software to verify and deduct their mass spectrometry information.This study laid an important foundation for the basic research of WDD pharmacodynamic substances.Three,two weeks of drug intervention in MetS rats by WDD and metformin,respectively,were used to compare the changes of serum biochemical indicators and the expression of metabolic nuclear receptors in liver tissues to further reveal the relationship between WDD and metabolic nuclear receptors.It was observed that WDD significantly improved MetS symptoms in rats,and the effect was significantly higher than metformin,which confirmed that WDD was effective drug for MetS treatment.After WDD intervention,the expression levels of PPAR?,PPAR? and LXR? in the livers of MetS rats were significantly up-regulated(P<0.05),which confirmed the molecular target results of network pharmacology research in WDD.Four,the detection of serum ABCA1 expression in rats,QSAR modeling,molecular docking and UHPLC-Q Exactive Orbitrap/MS technology were combined to study the pharmacological organic components and mechanism of WDD against MetS by upregulation of HDL-C levels.It was revealed that two high-accuracy QSAR classification models for upregulating ABCA1 activity were established,the mechanism of WDD upregulating HDL-C expression was by acting on the LXR?/ABCA1 signaling pathway and the structures of 9 organic compounds in WDD targeting LXR? receptor and up-regulating ABCA1 activity were inferred,which provided an effective method for the discovery of new highly potent ABCA1 expression upregulators from the chemical constituents of traditional Chinese herbs.Five,the metabolomics methods(GC-TOF/MS),multivariate statistical techniques(PCA and PLS-DA),KEGG biological pathway analysis were integrated to clarify the pharmacodynamic mechanism of WDD against MetS from the overall level of endogenous small molecular metabolites.It was concluded that MetS caused significant changes in 39 small molecular metabolites and 7 metabolic pathways in vivo(P<0.05),which can be identified as potential biomarkers of MetS.Also,six metabolic pathways were significantly affected by two weeks of WDD intervention in MetS rats(P<0.05).Then,the mechanism of WDD inducing changes of differential metabolite levels was explored by the chemical information method combined with UHPLC-Q Exactive Orbitrap/MS technology,which led to qualitatively structural identification of 9 glycosides acting on multi-metabolic enzymes in WDD. |
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