| Objective:Based on iTRAQ proteomi c research technology,this experiment takes the 16 month old mice as the experimental object,through the comparative analysis of the changes of protein in the liver mitochondria of the 3-month-old and 16 month old female and male mice,excavates the protein markers closely related to aging,and explores the influence of Sijunzi Decoction on the biological markers of aging related protein,so as to explain the aging from the perspective of proteomics The mechanism of the occurrence and development of aging and the biological basis of Sijunzi Decoction interfering with the age-related syndrome of mice provide scientific basis for the study of the relationship between formula and syndrome.Methods:Based on the previous study of this research group,10 male and 10 female SPF clean ICR mice of 3 months old were selected as the blank group,20 male and 20 female ICR mice of 16 months old were randomly divided into Sijunzi Decoction group and model group,10 male and 10 female mice in each group.Sijunzi Decoction group was administrated with concentrated solution(the dosage was about 9.75g/kg·d),and the other groups were administrated with normal saline of the same volume ratio.On the 3 1st day after administration,the mice were killed by cervical dislocation method and their livers were removed.The samples were labeled and frozen with liquid nitrogen,and immediately stored in-80℃ refrigerator for future use.The changes of biomarkers of mitochondrial protein in hepatocytes of natural aging mice were studied by relative and absolute quantitative techniques of isotope labeling.Results:1.Based on the analysis of proteomic iTRAQ technology,it was found that there were protein biomarkers related to aging in the mitochondria of liver cells of model mice.After the intervention of Sijunzi Decoction,there were some retrogressions,including:UDP glucuronosyltransferase,collagen alpha-5(VI)chain,integrin alpha-9(integrin)Alpha-9,non-specific serine/threonine protein kinase,hyaluronidase,guanine nucleotide exchange factor Vav2,40S ribosomal protein S 1 5,cytochrome P450 2j 6,etc2.Analyze the protein markers interfered by Sijunzi Decoction by gene ontology,and find that the intervention of Sijunzi Decoction on the aging related protein markers is mainly reflected in protein transport,steroid metabolic process,ribosome decomposition It includes:disassembly,autophagy,lysosome,mitochondrion,ribosome,endoplasmic reticulum,hydrolase activity,catalytic activity,and transfer activity3.After enrichment analysis of protein markers through KEGG pathway pathway pathway,it was found that the intervention of Sijunzi Decoction on aging related proteins was mainly related to metabolic pathways,endocy tosi s,focal adhesion s,splice,Alzheimer’s disease and Huntington’s disease Disease,ECM receptor interaction,ribosome,hypertrophic cardiomyopathy(HCM),apoptosis,mTOR signaling pathway and so onConclusion:1.Compared with the 3-month-old blank mice,the protein markers in the liver samples of the 16-month-old model mice were significantly different,indicating that the natural aging can cause the changes of protein markers and related biological pathways.2.Through the study of proteomics,we found that Sijunzi Decoction can regulate the metabolic pathway,endocytosis,adhesion spot,spliceosome,Alzheimer’s disease,Huntington’s disease,ECM receptor interaction,ribosome,hypertrophic cardiomyopathy(HCM),apoptosis,mTOR signaling pathway and so on.By regulating the related proteins of these signaling pathways,it can play a protective and regulatory role in the aging related dysfunction of the body3.Through go analysis of protein markers,it was found that Sijunzi Decoction mainly regulated protein transport,steroid metabolism,ribosome decomposition,autophagy and other biological processes,affected important cell components such as lysosome,endoplasmic reticulum,mitochondria,ribosome,and participated in the regulation of molecular physiological functions such as hydrolase activity,transferase activity and catalytic activity. |
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