Proteomics Study On The Intervention Effect Of Siwu Tang On 16 Month Old Mice

Objective:By using high-throughput iTRAQ proteomics technology,the changes of mitochondrial protein in the liver of 3-month-old and 16-month-old female and male mice were compared,and the mechanism of Si wu Decoction regulating age-related protein in 16-month-old mice was analyzed.The biological basis of Si wu Decoction interfering age-related syndrome was further analyzed based on database and literature data,so as to comprehensively analyze the mechanism of aging,To provide new theoretical basis and data support for the development of TCM Prevention and treatment measuresMethod:Ten ICR female and male mice at the age of 3 months were selected as the young control group;30 ICR female and male mice at the age of 16 months were randomly divided into Siwutang group,vitamin E group and blank control group.Siwutang group and vitamin E group were administrated by gavage,while young control group and blank control group were administrated by gavage with normal saline of the same volume ratio.The mice were sacrificed on the 3 1st day after administration,the livers were taken out,and the liquid nitrogen was quickly frozen and immediately transferred to a refrigerator at minus 80 ℃ for storage.Hepatic mitochondria were extracted,based on proteomics research methods,based on iTRAQ technology,combined with LC-MS/MS mass spectrometry,and screened for differential proteins through the UniProt release(2016-07)database.GO analysis and KEGG were performed on the differential proteins in each group pathway analysis.Intersect the differential proteins of each group to further screen protein markers,study the mitochondrial protein biomarkers of hepatocytes in natural aging model mice,and analyze the protein biomarkers of Siwutang intervention for age-related diseases and syndromes.To determine the protein biomarkers and pathways that have the highest correlation with the intervention of Siwu Tang,and use proteomic data to explore the biological basis of Siwu Tang’s intervention in natural aging mice.Result:1.Proteomics analysis and identification of mitochondrial protein biomarkers in hepatocytes of natural aging model mice by iTRAQ proteomics,and determination of related protein biomarkers during aging of mice treated by Siwu Tang.Including Peripheral plasma membrane protein CASK,Immunoglobulin heavy variable 1-81,ADP-sugar pyrophosphatase,Protein phosphatase 1G,Cellular nucleic acid-binding protein,Gly cerate kinase,Ubiquitin carboxyl-terminal hydrolase isozyme L5,Hy aluronidase,Actin-related protein 2/3 complex subunit 1A,Serine/threonine-protein phosphatase,etc.2.The biological effects of Siwu Tang on biomarkers were analyzed by GO.The intervention of Siwu Tang on the biological process of protein markers in mice during aging is reflected in transportation,protein transport,cell cycle,cell adhesion,translation,nervous system development,Positive regulation of GTPase activity,cardiac muscle contraction,protein dephosphorylation,response to glucose,etc.Intervention of protein marker molecular function is reflected in protein binding,poly(A)RNA binding,nucleotide binding,hydrolase activity,RNA binding,protein homo dimerization activity,catalytic activity,actin binding,protein kinase binding,Protein complex binding,actin filament binding,transport activity,lipid binding,cytoskeleton protein binding,phosphoprotein phosphatase activity,oxidoreductase activity,etc.3.From the analysis of KEGG pathway,the intervention effect of Siwu Tang on related protein markers in mouse aging and RNA transport,focal adhesions,regulation of actin cytoskeleton,hypertrophic cardiomyopathy(HCM),dilated cardiomyopathy,insulin signaling pathway and so onConclusion:1.Comparing the 16-month-old blank group with the 3-month-old young control group mice,significant differences in protein markers in liver mitochondrial samples were found,suggesting that natural ageing may cause changes in mouse protein-related biological pathways2.Siwu Decoction has a callback effect on the protein markers in liver mitochondria samples of 16-month-old female and male mice.The regulated differential proteins are mainly enrichedin RNA transport,focal adhesions,regulation of actin cy toskeleton,hypertrophic cardiomyopathy(HCM),dilated cardiomyopathy,insulin signaling pathway,etc.It is suggested that Siwu Tang can regulate the changes of various pathway proteins caused by aging,improve the related biological changes caused by aging,and regulate the related body dysfunction caused by aging.The regulation effect of vitamin E on protein level in 16-month-old mice is not obvious.