| Objective:In this study,nuclear magnetic metabonomics analysis was used to establish insulin resistance(IR)rat pancreatic tissue metabolic fingerprints and screening of metabolic markers,combined with previous serum and urine metabolomics results for comprehensive analysis,to find serum,urine and Differential metabolites and pathways associated with exercise intervention IR in pancreatic tissue,use of biomolecular networks to build relevant metabolic and disease networks,correlate target predictions with metabolomics results,and explore improvement of exercise intervention from the perspective of metabolic network regulation The mechanism of action of IR.Methods:In this study,~1H-NMR was performed on the pancreatic tissue of IR rats.The nuclear magnetic spectrum was calibrated,phase and baseline calibrated,integrated,and normalized using MestReNova(version 8.0.1,Mestrelab Research,Santiago de Compostella,Spain)software.Multivariate statistical analysis was performed using the software SIMCA-P 13.0 and combined.Corresponding S-plot,VIP values and t-test results,looking for endogenous differential metabolites and related biological metabolic pathways related to exercise intervention IR,and its biological nature relying on the biological information database KEGG and related literature research results to achieve further mining,and evaluate the diagnostic predictive value of endogenous differential metabolites by ROC curve analysis.Finally,by integrating the results of ~1H-NMR metabolomics of pre-laboratory serum and urine,the potential biomarkers related to exercise intervention IR and the high metabolic pathways were screened.Combined with the method of biomolecule network,the key to the pathway was selected.Integration of genes with disease genes,combining metabolites,metabolic pathways,IR disease genes,and IR diseases to explore the pathogenesis of high-fat-induced IR and the mechanism of action intervention IR.Results:(1)Analysis of N,IR,and IRe in pancreatic tissue of high-fat diet IR rats by ~1H-NMR metabolomic analysis revealed amino acids,organic acids,choline,inositol,inosine,and phosphoric acid Eighteen endogenous metabolites such as choline were selected and 11 IR-related differential metabolites were screened out.The ROC curve predicts five potential biomarkers related to exercise intervention IR.(2)Obtained disease-related gene information through database search and text mining of GeneCards,NCBI,and CooLGeN.The constructed IR disease network contains a total of536 nodes and 8237 edges,and performs topology and module analysis on the disease network,according to the Degree value.The top 50 genes were listed and 8 disease modules were obtained.The biologic annotation of the genes in the IR disease module and the KEGG pathway enrichment analysis were performed by DAVID.The biological processes marked by GO annotation were cell proliferation,apoptosis,cytokine response,lipid metabolism,and glucose metabolism.The results of the analysis showed that the biological pathways that are significantly enriched include HIF-1,TNF,PPAR,PI3K-Akt,AMPK,mTOR,and insulin signaling pathways.(3)Thirty-three IR-related differential metabolites and 23 differential metabolites related to exercise intervention IR detected in serum,urine,and pancreas based on ~1H-NMR metabolomics were analyzed and screened separately.6important pathways.By analyzing the relationship between the genes on the pathway and the IR disease genes,four pathways with direct relationships were found.Proteins encoding PCK1,GFPT2,GPT,GOT2,CAT,GGT1,and GAD2 are key genes in the IR pathway,and CAT,PCK1,GGT1,and GAD2are key genes for exercise intervention.Conclusion:The major metabolic pathways involved in exercise intervention IR include glyoxylate and dicarboxylic acid metabolism,tricarboxylic acid cycle,alanine,aspartate and glutamate metabolism,and taurine and hypotaurine metabolism.The key pathway genes for exercise intervention IR are CAT,PCK1,GGT1,and GAD2.Exercise may improve IR status by regulating the expression of these genes.The internal and external integration analysis of metabonomics results has important implications for the systematic analysis of IR pathogenesis and the mechanism of action intervention IR. |
