A Biomarker Study Of Hyperuricemia Based On Untargeted And Targeted Metabolomics

Hyperuricemia(HUA)has become the "fourth-highest" after hypertension,hyperlipidemia,and hyperglycemia.It seriously harms human health and has become a public health problem that needs to resolve urgently.HUA is a complex metabolic syndrome with complex influencing factors.At present,the exact etiology and pathogenesis of HUA are not fully understood.Therefore,it is still of great significance to find effective biomarkers and explore the pathogenesis of HUA.Metabolomics reflects the influence of internal and external factors on system metabolism,explains the changes in metabolite levels during the development of diseases,and reveals the molecular mechanism of pathogenesis.It can provide a basis for searching for HUA biomarkers and exploring metabolic disorders in the occurrence and development of HUA.It is helpful to understand HUA in depth and clarify the exact etiology and mechanism of HUA.Metabolomics is divided into untargeted metabolomics and targeted metabolomics according to different research modes.Both have their advantages and disadvantages.Each other's advantages can fully utilize by combining the two so that the results of metabolomics research can be perfect.At present,most metabolomics studies on HUA only carry out untargeted metabolomics studies.But there are insurmountable difficulties.In the present study,untargeted and targeted metabolomics approaches were performed to screen and verify the biomarkers of HUA patients.And then further explore the occurrence and development of HUA.The main research contents of this paper are as follows:1.Data analysis of untargeted serum metabolomics in HUAAccording to the previous comparison results of the sample pretreatment methods obtained by the research group,we extracted the data of serum samples processed respectively by MeOH/MeCN(1:9,v/v)and MeOH/MeCN(5:5,v/v)and detected by the UPLC-Q-TOF/MS analysis platform.After obtaining the data,untargeted metabolomics analysis was performed:the data was preprocessed to obtain a two-dimensional data matrix,perform multivariate statistical analysis,and screen and identify differential metabolites based on multivariate and univariate analysis and public databases,and perform metabolic pathway analysis based on differential metabolites,and candidate biomarkers were screened based on literature research and ROC Curve analysis.Results:70 different metabolites were screened and identified.The 7 metabolic pathways with the most significant changes in differential metabolites.And 12 candidate biomarkers were screened finally for subsequent verification.2.Targeted serum metabolomics study of HUA based on UPLC-TQ-MSThe twelve candidate biomarkers were divided into four polar metabolites and eight lipid metabolites according to their physical and chemical characteristics.Based on the UPLC-TQ-MS analysis platform,with a simulated serum sample added with mixed standards as the research object,the targeted metabolomics detection methods of serum polar metabolites and lipids metabolites were established respectively,and methodological verifications were conducted respectively.The methodological results showed that the two established methods have great specificity.The standard curves have good linearity in the linear range.The intra-and inter-day precision and accuracy,the lower limit of quantification,extraction recovery,matrix effect,and stability meet the requirements of biological sample analysis.The two established methods are suitable for the determination of candidate biomarkers.Afterwards the established method was applied to patients' serum samples(20 HUA patients and 20 normal people)to determine the content of polar metabolites and lipid metabolites.Assay results showed that the contents of L-Tyrosine,L-Phenylalanine,Arachidonic acid,Linoleic acid,Oleic acid,Stearic acid,LysoPC(18:0),LysoPC(18:1(9Z)),and LysoPC(16:0)in the HUA serum significantly lower than the contents in normal serum to varying degrees,but the contents of L-Valine,L-Lactic acid,and Palmitic acid significantly higher than the contents in normal serum to varying degrees.3.Investigation of pathogenesis of hyperuricemia based on untargeted and targeted metabolomicsCombined with the results of untargeted and targeted metabolomics.For 9 biomarkers with the same content concentration change trend in untargeted and targeted metabolomics(L-Tyrosine,L-Phenylalanine,Arachidonic acid,Linoleic acid,Oleic acid,Stearic acid,LysoPC(18:0),LysoPC(18:1(9Z))and LysoPC(16:0)),the reliabilities were determined firstly.Then metabolic characteristics of each substance were analyzed,metabolic functions and activities with physiological phenomena were linked,the changes within the HUA body and possible mechanisms of occurrence and development were explored.For 3 biomarkers with the inconsistent content concentration change trend in untargeted and targeted metabolomics(L-Valine,L-Lactic acid and Palmitic acid),the targeted metabolomics results were considered more reliable.According to the results of targeted metabolomics experiments,the characteristics and activities of metabolism were analyzed,and the reliability was verified.