| Objective:Metabonomics is a new method of studying dynamic change of endogenousmetabolite in biosystem and has been increasingly used as an effective tool for diseasediagnosis to elucidate significant changes in metabolism, also an ideal tool for studyingtraditional medicine theories.In our study, With the help of the recent advent oftechnologies for quantitative and comprehensive metabolites analyses, which provides amore sensitive and specific diagnosis than single biomarker approaches, and revealdistinct differences in metabolism between diseased individuals and healthy ones in anovel systems-level.Methods:1．In the present study, NMR was used in the urine metabolomics study, verifyingthe high performance of the system in separating endogenous metabolites and its abilityas a nearly diagnostic method for uygur medicine.2．Through studying the metabolic variations in urine of type2diabetes mellituspatients and healthy volunteers with UPLC-TOF-MS, more disease information could begain to help finding the common and specific biomarkers among the diseases, which gavenew eyes on the understanding of the mechanism of the diseases.Here, T2DM andmetabonomics were investigated by comparing small molecules in urine samples fromT2DM patients and healthy controls, the potential biomarkers of DM were found. 3．A robust and sensitive liquid chromatography spectrometry (HPLC) method wasdeveloped and optimized for quantification of various amino acids in serum and urine.Then the data was imported through the analysis by principal component analysis (PCA),and the least squares discriminant analysis (PLS-DA). The analytical serum amino acidsplatform can applied to investigate the effect of diabetes on amino acids in serum, urineand diabetes rats, Then, results shows the classification accuracy were good.4．To investigate the serum free fatty acid variation in diabetic patients and heathyvolunteers. the human serum were methylesterifiled by H2SO4/CH3OH, extracted byn-hexane, FID detector,7kinds of fatty acids can be separated completely with a goodlinear and chemometric resolution method were used to discriminated diabetes patientsand controls.Results:1．The application of NMR detection has been investigated for the production ofglobal metabolite profiles from human urine, Chemomatrics was performed to detect theperturbation metabolites as many as possible. Furthermore, the potential biomarkers werescreened out, which might be the target components in the future pathogenesis research,as well as predicted model was builded up using PLS-DA. The metabonomics basedNMR and OPLS can prove the difference in differentiated traditional Uygur Medicinesyndrome with diabetes, Eatabilished the approach with NMR for metabonomic study,some specific urine endogenous metabolites changes in the metabolic composition ofurine samples from diabetes, there are Elevated glucose,hippuric acid, betaine, taurine,etc.; leucine, isoleucine, creatinine, citric acid, dimethylamine, and other content is lowerthan normal control.2．A UPLC Q-TOF MS system was employed to distinguish the urine global profilesof38type2diabetes mellitus patients and32heatlty volunteers. PCA and PLS-DA wasused for group differentiation and potential biomarkers selection. As shown in the scoresplot, the distinct clustering among diabetes mellitus patients and health controls wasobserved. In carbohydrate metabolites, sodium glucuronic acid content was significantlyhigher than the normal control group, compared with the normal control group, thesubstance of the fatty acids were higher than control group,20-carboxy-arachidonic acid,palmitic acid in the urine of patients with diabetes were higher than in normal subjects.Metabolites of amino acids such as tyrosine, N-beta-alanyl-L histidine, tryptophancontent is lower than normal, glutaric acid, nicotinoyloxy glycine, aspartic acid, phenylalanine, etc. content were increased than normal people. These metabolites areassociated with in glucose and lipid regulating substances.This study indicatedthepotential of metabolomic strategies for explanation of the mechanism of diseases anddetection of specifical biomarkers.3．The HPLC based serum and urine amino acid metabolomics approach was alsoused to investigate the pathophysiological variation among type2diabetes mellituspatients compared to health control. The derivatization condition was optimized. Patternrecognition was carried out with PCA and PLS-DA.Then, the serum of type2diabetesrats were analysis by this method.In diabetes, the amino acid metabolism includingaspartate,glutamic, serine, threonine were found as the most obvious changed aminoacid.although aspartic acid and phenylalanine were significantly higher than normal by ttest, a significant decline in the amino acid are alanine, glycine and lysine. Metaboliteprofiles in the urine was analysised in a multivariate analysis and the result shows thatserine, threonine, taurine were most contributed to inter-group, one-dimensionalstatistical analysis found that the urine taurine, arginine, serine and histidine significantlylower than normal, and glutamic acid were increased significantly. Are two ways to dealwith the results obtained can be complementary.4．The GC based serum metabolomics approach was also used to investigate thepathophysiological variation among type2diabetes mellitus patients compared to healthcontrol. The derivatization condition was optimized. Pattern recognition was carried outwith PCA and OPLS-DA were used to analysis All fatty acids were suceessfullyidentified and quantified, and stable plasma FFA metabolie Profiling of both the controlsand TZDM patients. The results found elevated Myristicacid in the serum of withdiabetes patients, while oleic acid in diabetic patients specifically down regulatedcompared with healthy people, which suggested that these two substances were potentialspecific markers to distinguish between diabetes and control and fatty acid metabolic canprovide important experimental basis for diagonosis.Conclusion:The NMR separation of urine samples was performed and UPLC-TOF-MSmetabolomics study can clearly classify the diabetes groups and control group. Themetabolites gotton from NMR were different from those from UPLC/MS, but thedifferent metabolites maily including amino acid, glucose, and organic acid. Comparedwith healthy controls, the glycolysis metabolism, and amino acid metabolism of diabetes mellitus patients were disordered to some degree.It indicated that both common andspecific metabolic pathways of diabetes mellitus were disordered. The relationshipbetween the two methods could b efurther studied according to the results of the presenttest.The HPLC and GC based serum and urine metabolomics approach was also used toinvestigate the pathophysiological variation among type2diabetes mellitus patients. Thederivatization condition was optimized. Pattern recognition was carried out with PCA andPLS-DA to get the similar results. This study would provide important significance fordiagnosis and evaluation of the therapeutic efficacy for diabctie mellitus. |
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