| ABJECTIVEThe metabolic profile and biomarkers of the blood,spleen and kidney tissues of the natural aging model mice were studied by metabonomics,and the effects of Sijunzi decoction on the metabolic profile and biomarkers of the natural aging model mice were analyzed.On the basis of iTRAQ technology,the protein differential expression profiles related to natural senescence were compared and identified by the method of proteomics,and the protein markers of senescence were found,and the modulation of Sijunzi decoction on the biomarkers of aging related protein was studied.Combined with the data of metabonomics and proteomics analysis,the biological pathway with the highest correlation with the intervention of Sijunzi decoction was analyzed,which provided the basis for the biological basis of the intervention of Sijunzi decoction to the syndrome of natural aging related diseases.METHODBased on the preliminary study of the experimental group,the preparation and UPLC-HDMS analysis methods were established and applied to the samples of mice's blood,spleen and kidney tissue.The UPLC-HDMS technology platform was used to evaluate the effect of Sijunzi decoction on the intervention of natural aging model mice,and the effect of the Sijunzi decoction on the blood and spleen tissue was studied.The influence of renal tissue metabolic profile and biomarkers.On the basis of the natural aging model,the metabolic group evaluation of the intervention of Sijunzi decoction was carried out to study the influence of Sijunzi decoction on the metabolic profile of blood,spleen and kidney tissue and biomarkers.Further based on the natural aging model mice,the hepatocyte mitochondria were analyzed by proteomics,based on iTRAQ technology,combined with LC-MS/MS mass spectrometry,combined with GO annotation and KEGG pathway analysis,to study the mitochondrial protein biomarkers of the liver cells of the natural aging model mice,and to intervene the aging phase of Sijunzi decoction.According to the analysis of disease protein biomarkers,we determined the highest protein biomarkers and pathways related to the intervention of Sijunzi decoction.Finally,based on the data of metabonomics and proteomics,the mechanism of the intervention of Sijunzi decoction on the damage of the blood,spleen,kidney and hepatocyte mitochondria in the mice of natural aging model was investigated.RESULT1.based on the metabonomics technique,the effects of the intervention of Sijunzi decoction on the metabolic profile and biomarkers of the natural aging model mice were studied.Compared with the model group,the metabolic profiles of the blood,spleen and kidney tissues of Sijunzi decoction were obviously changed.The results of the PCA analysis of the metabolic profiles of the 3 biological samples showed that the results of the metabolic profiles of these 3 biological samples showed that Sijunzi decoction group was close to the blank group and kept away from the model group,and returned to the normal group.The identified blood biomarkers related to natural aging were analyzed.Among the 25 biomarkers of the male model blood samples,22 of them were adjusted,19 of them were statistically significant,and among the 26 biomarkers of the female model blood samples,14 of them were adjusted.5 of them have statistical significance.The biomarkers are mainly related to the Cysteine and methionine metabolism,Steroid biosynthesis,Arginine and proline metabolism and other pathways.The identified spleen tissue biomarkers related to natural aging were analyzed.Among the 26 biomarkers of the male model spleen tissue samples,24 of them were adjusted,21 of them were statistically significant,and the female model spleen tissue samples were of 26 biomarkers,and 22 of them were given to the drug group.11 of them have statistical significance.The biomarkers are mainly related to the D-Glutamine and D-glutamate metabolism,Phenylalanine,tyrosine and tryptophan biosynthesis,Taurine and hypotaurine metabolism,Glutathione metabolism,Arachidonic acid metabolism,Alanine,aspartate and glutamate metabolism,Pentose and glucuronate interconversions and other pathways.The identified biomarkers related to natural aging were analyzed.Among the 23 biomarkers of the male model kidney tissue,18 of them were adjusted,13 of them were statistically significant,and among the 24 biomarkers of the female model kidney tissue,23 of them were given to the drug group.11 of them have statistical significance.The biomarkers are mainly related to D-Glutamine and D-glutamate metabolism?Arachidonic acid metabolism?Biotin metabolism?Taurine and hypotaurine metabolism?Alanine,aspartate and glutamate metabolism and other pathways.2.iTRAQ protein group was used to analyze and identify the white biomarkers of egg hepatocytes in the natural aging model mice,and to determine the biomarkers of the aging related protein biomarkers for the intervention of Sijunzi decoction.The intervening effect of the Sijunzi decoction on biomarkers was analyzed by GO.The intervention effect of Sijunzi decoction on the markers of aging related protein was reflected in oxidation-reduction process,lipid metabolic process,metabolic process,fatty acid metabolic process,protein transport,fatty acid transport,mitochondrion,oxidoreductase activity,hydrolase activity,nucleotide binding,protein binding,transferase activity,immune system process,mitochondrion,protein homodimerization activity,structural molecule activity,catalytic activity.According to the KEGG pathway analysis,the intervention of Sijunzi decoction to the aging related protein markers is related to Metabolic pathways,Peroxisome,PPAR signaling pathway,Carbon metabolism,Alzheimer's disease,Non-alcoholic fatty liver disease,Parkinson's disease,Insulin signaling pathway.CONCLUSIONMetabonomics studies found that Sijunzi decoction plays an important role in the metabolic process of amino acids.By regulating the metabolism of cysteine and methionine,arginine and proline metabolism,D-glutamine and glutamate metabolism,taurine and taurine metabolism,glutathione metabolism,and biological Sudache,the increase of age in mice is improved.Related metabolic disorders and free radical damage.The study of proteomics found that Sijunzi decoction was redox reaction,lipid metabolism,metabolic process,fatty acid metabolism,protein transport,fatty acid transport,oxidoreductase,nucleotides binding,protein binding,immune system process,mitochondrial metabolic pathway,peroxisome,Alzheimer's disease and so on.The approach has a regulatory role.By regulating the related proteins of these signaling pathways,it can play a protective and regulatory role in the related functional disorders of aging.Combined with metabonomics and proteomics for multi sample and high throughput analysis,it was found that Sijunzi decoction had protective effects on amino acid metabolism,immune system and lipid metabolism,protein transport and mitochondria.It is indicated that the intervention effect of Sijunzi decoction on the related diseases of aging is achieved through these biological pathways. |
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