The Biological Basis Of Deficiency And Excess Syndrome In Rheumatoid Arthritis Based On Text Mining And Metabolomics

Background:Rheumatoid arthritis(RA),which belongs to the category of "Bi Zheng" in traditional Chinese medicine(TCM),is the dominant disease in clinical treatment of TCM.Syndrome differentiation and classification of RA is the premise of determining the corresponding treatment principles and methods,which reflects the theoretical characteristics and advantages of syndrome differentiation and treatment of RA in TCM diagnosis and treatment.Deficiency and excess syndrome,as the basic types of eight-principle syndrome differentiation,are one of the diagnostic elements that must be identified in the treatment of RA syndrome differentiation.For a long time,there has been a lack of objective quantitative standards in the study of TCM syndrome classification.How to use modern scientific methods to reveal the scientific connotation contained in TCM syndrome theory,explore the biological basis of TCM syndrome classification,and clarify the micro mechanism of TCM syndrome differentiation and treatment is very important.Deficiency of liver and kidney syndrome and cold-dampness obstruction syndrome are the main types in the classification of deficiency and excess syndrome of RA.Selecting them as starting points for the preliminary exploration of deficiency and excess syndrome of RA will help to provide reliable scientific basis,technical and theoretical support for more comprehensive and systematic study of TCM syndrome classification in the future.What's more,it can promote the development and inheritance of TCM syndrome theory,and further guide the more accurate treatment based on syndrome differentiation in clinical RA to obtain better clinical efficacy.Objective:The purpose of this study is to explore the biological characteristics of deficiency and excess syndrome of RA through text mining technology combined with bioinformatics analysis.At the same time,the serum samples of patients with RA liver and kidney deficiency syndrome and cold-dampness syndrome were studied by metabonomics,and the biological characteristics of RA liver-kidney deficiency syndrome and cold-dampness syndrome were obtained,and compared with the text mining results of deficiency and excess syndrome of RA,to obtain reliable RA empirical evidence on biological basis.Methods:1.Research on text mining of deficiency and excess syndrome of RA:Conduct text retrieval using text mining technology based on PubMed,SinoMed database,with "Lei feng shi guan jie yan","Rheumatoid Arthritis","Arthritis,Rheumatoid","Deficiency syndrome","Deficiency pattern","Xu Zheng","Excess syndrome","Excess pattern","Shi Zheng" and other topic words.After data cleaning,processing,visualization,and bioinformatics analysis combined with Ingenuity Pathway Analysis(IPA)online bioinformation software platform,the biological characteristics of deficiency and excess syndrome of RA wereobtained.2.Metabonomics study of RA liver and kidney deficiency syndrome and cold-dampness syndrome:(1)Clinical research subjects and serum samples were collected and preserved according to the established diagnostic criteria of RA,TCM syndrome diagnostic criteria,inclusion criteria,exclusion criteria.(2)The serum samples were pretreated and metabonomics was detected by UPLC-QTOF-MS technology.After the original data were processed,multivariate statistical analysis and identification of differential metabolites were carried out,and bioinformatics analysis was carried out combined with IPA to obtain the biological characteristics of RA liver and kidney deficiency syndrome and cold-dampness obstruction syndrome at metabolic level,which were further compared with the text miningresults of deficiency and excess syndrome of TCM.Results:1.Through the research on text mining of deficiency and excess syndrome of TCM,633 search results of deficiency syndrome and 208 excess syndrome search results were obtained in PubMed and SinoMed databases(information up to December 31,2019).By comparing the one-dimensional results of virtual and empirical text mining related biomolecules,it is shown that the frequency of biomolecules related to deficiency syndrome is higher than that of excess syndrome,there are 14 biomolecules related to deficiency syndrome whose frequency are more than 30:IgG,Nitric Oxide,CD4,Glutl,IgE,CD18,Dopamine,SOD,Testosterone,IL-2,IgM,5-hydroxytryptamine,Cholesterol and INF-?;However,there are only 2 biomolecules of excess syndrome whose frequency is more than 30:Nitric Oxide and Mineralocorticoid.Among the top 15 biomolecules,there are four identical biomolecules:IgG,Nitric Oxide,IgE,CRP.Text mining extracted 416 related biomolecule combinations of deficiency syndrome and 276 related biomolecule combinations of empirical evidence.In deficiency syndrome,IgG and IgM are the most closely related,followed by CD4 and CD8,CD18 and Leukocytes.In the excess syndrome,Mineralocorticoid and Hydroxysteroid Dehydrogenases are the most closely related,followed by Renin and Aldosterone,Hydrocortisone and aldosterone.2.After IPA analysis,the unique biological pathways of biomolecules related to RA deficiency syndrome were obtained(top 5):mismatch repair of eukaryotes,ovarian cancer signaling,regulation of the epithelial-mesenchymal transition pathway,pancreatic adenocarcinoma signaling and hereditary breast cancer signaling.The specific biological pathways of biomolecules related to RA excess syndrome(top 5)are:hematopoiesis from pluripotent stem cells,Thl pathway,HMGB1 signaling,Thl and Th2 activation pathway and role of cytokines in mediating communication between immune cells.Eight functions and diseases closely related to biomolecules related to RA deficiency syndrome were obtained:metabolic disease,cell death and survival,nutritional disease,cellular movement,molecular transport,vitamin and mineral metabolism,hereditary disorder and psychological disorders.Eight functions and diseases closely related to RA excess syndrome were obtained:cardiovascular disease,cellular function and maintenance,developmental disorder,cellular growth and proliferation,neurological disease,protein synthesis,hypersensitivity response and cellular assembly and organization.3.This study included 19 female patients with RA liver and kidney deficiency syndrome,41 female patients with cold-dampness syndrome and 30 healthy women.Through the metabonomic study of serum samples,25 differential metabolites of RA liver and kidney deficiency syndrome and 24 differential metabolites of RA cold-dampness obstruction syndrome were obtained.Among them,the two syndromes contain 19 different metabolites:L-Homocysteicacid,LysoPE(P-16:0/0:0)),S-Nitrosoglutathione,5-Thymidylic acid,SN38 glucuronide,PE(22:0/24:0),PC(24:0/24:1(15Z)),Pyruvic acid,D-Ribose,Gamma-Glutamylserine,PE(22:0/24:1(15Z)),Inosinic acid,N(omega)-Hydroxy arginine,LysoPC(16:0/0:0),Bisdiphosphoinositol tetrakisphosphate,Citric acid,Octanoyl-CoA,LysoPC(P-18:0/0:0),PC(O-22:1(13Z)/22:3(10Z,13Z,16Z)).There are 6 differential metabolites peculiar to RA with liver and kidney deficiency syndrome:5-Methoxytryptamine,4-Fumarylacetoacetic acid,LysoPC(1 8:1(9Z)/0:0),LysoPC(20:1(11Z)/0:0),Glucosylceramide(d18:1/12:0),PE(20:3(5Z,8Z,11Z)/22:6(4Z,7Z,10Z,13Z,16Z,19Z)).There are 5 differential metabolites peculiar to RA with cold-dampness obstruction syndrome:Gamma-Aminobutyric acid,LysoPA(18:1(9Z)/0:0),DG(18:3(6Z,9Z,12Z)/22:6(4Z,7Z,10Z,13Z,16Z,19Z)/0:0),L-Targinine,PC(24:0/24:0).4.After IPA analysis,19 differential metabolites related biological pathways were obtained in RA liver and kidney deficiency syndrome and cold-dampness obstruction syndrome(top 5):L-cysteine Degradation ?,Glycine Biosynthesis ?,Alanine Degradation ?,Alanine Biosynthesis ? and L-serine Degradation.Molecular functions and diseases related to two syndromes were obtained(top 5):Immunological Disease,Small Molecule Biochemistry,Cell Morphology,Cell Cycle,Connective tissue disorders.3 biological pathways related to the specific differential metabolites of RA liver and kidney deficiency syndrome were obtained:Melatonin Degradation ?,Tyrosine Degradation I,Superpathway of Melatonin Degradation.6 molecular functions related to RA liver and kidney deficiency syndrome were obtained:Amino acid metabolism,Cell-To-Cell signaling and interaction,Inflammatory response,organismal injury and abnormalities,molecular transport and nucleic acid metabolism.5 biological pathways related to different metabolites of RA cold-dampness syndrome were obtained:Glutamate dependent acid resistance,4-aminobutyrate degradation ?,glutamate degradation ?(via 4-aminobutyrate),putrescine degradation ?,GABA receptor signaling and neuroinflammation signaling pathway.8 molecular functions related to cold-dampness obstruction syndrome were obtained:cellular growth and proliferation,cellular assembly and organization,cellular compromise,cellular development,DNA replication,recombination and repair,energy production,post-translational modification and free radical scavenging.5.The results of metabonomics research and text mining research were compared.The results showed that among the molecular functions related to differential metabolites of RA liver and kidney deficiency syndrome and text mining RA deficiency syndrome,molecular transport was common to both,and the molecule of differential metabolites of RA liver and kidney deficiency syndrome in this function was 5-methoxytryptamine.The biomolecules related to RA deficiency syndrome are ADC Y,cyclic AMP,hydrogen peroxide,melatonin,BDNF,CCR5,CD4,CD40,CRP,CXCR4,DRD2,DRD4,HTR4,IL2,IL6,ITGB2,JAK2,KCNJ11,PDE5A,SLC2A1,SMAD4,STAT3,TNF,VEGFA,VIP,AR,BAX and ADCY,and there are complex and close relationships between molecules.Among the molecular functions related to differential metabolites of RA cold-dampness obstruction syndrome and text mining RA excess syndrome,cellular growth and proliferation and cellular assembly and organization are in common.The molecules of differential metabolites of RA cold-dampness syndrome in cell growth and cellular growth and proliferation function are gamma-aminobutyric acid,LysoPA(18:1(9Z)/0:0)/monooleylphosphatidic acid and L-Targinine/N(G)-monomethyl-D-arginine.It is demonstrated that the related biomolecules in this function are IL2,IL4,IL6,PLAT,TNF,CD4,CD8A and CRP.The molecules of differential metabolites of RA cold-dampness syndrome in cellular assembly and organization function are gamma-aminobutyricacid and L-Targinine/N(G)-monomethyl-D-arginine,and the related biomolecules in this function are IL4,IL6,TNF and CRP.Conclusion:1.5-methoxytryptamine,a differential metabolite,mediates biological pathways of melatonin degradation ? and superpathway of melatonin degradation melatonin,and participates in molecular transport,which is the biological feature of RA liver and kidney deficiency syndrome in female patients.And it is also the potential biological basis of deficiency syndrome of RA.2.Differential metabolite gamma-aminobutyricacid/GABA mediates 4-aminobutyrate degradation ?,glutamate degradation ?(via4-aminobutyrate)and GABA Receptor Signaling biological pathways,and participates in cellular growth and proliferation and cellular assembly and organization are the biological characteristics of RA cold-damp obstruction syndrome in female patients.And it is also the potential biological basis of excess syndrome of RA.3.Based on text mining and metabonomics technology,combined with bioinformatics analysis,the comparative verification of the biological characteristics of RA liver-kidney deficiency syndrome and cold-dampness obstruction syndrome can not only provide evidence support for further experimental verification,but also provide methods and technical references for TCM syndromes,especially the classification of RA deficiency and excess syndromes.