Screening Of Aging-related Proteins By Moxibustion Therapy Of Mongolian Medicine

Objective: To explore the research on the influence of aging-related proteins based on TMT technology.Methods: 1.Thirty SD rats,male and female,were randomly divided into normal control group,model control group and Dinghui acupoint group according to their body weight,with 10 rats in each group.The model was established by subcutaneous injection molding of 5% D-galactose in the neck,and the mold was molded for 42 days.After the successful modeling,the moxibustion was started,and the moxibustion was performed once every other day for 21 days.Observed and detected changes in body weight,serum catalase(CAT)activity,malondialdehyde(MDA)content,physical strength,learning and memory and other related indicators for statistical analysis.Then select the moxibustion top acupoint group and the model group,each group of 10 SD rats,half male and half female.2.The collected two groups of serum were subjected to serum high-abundance protein and SDS-PAGE electrophoresis and FASP enzymatic analysis,pre-primary analysis,Date Base database alignment,peptide TMT labeling.The moxibustion top acupoint group was labeled as(126,127,128)and the model group was labeled as(129,130,131).3.Mass spectrometry was performed using a high resolution mass spectrometer Q Exactive,and the data was searched for the Uniprot protein database using the Proteome Discoverer 2.1 software and MASCOT 2.6 protein firming software with a FDR < 0.01 standard.The difference in selection has significant results for the expression difference multiples greater than 1.2 times(up and down)and P <0.05,and the bioinformatics analysis of differentially expressed proteins was performed using a GO platform,KEGG pathway and other database platforms.Results: 1.(1)Effect of moxibustion on body weight of model rats: There were significant differences between normal group and Dinghui acupoint group and model control group(P<0.05 or P<0.01).(2)Behavior of moxibustion on model rats Effects of academic indicators: 1 The effect of moxibustion on TRT: There were significant differences between the normal group and the top acupoint group and the model control group(P<0.01).The effect of moxibustion on EN: normal group and top acupoint group Compared with the model control group,there were significant differences(P<0.01).The effect of moxibustion on physical strength: There was significant difference between the normal group and the top acupoint group and the model control group(P<0.01).(3)Moxibustion treatment The effect of serum indexes in model rats: There were significant differences between the normal group and the top acupoint group compared with the model control group(P<0.01).2.Four differentially expressed proteins were screened,including fibrinogen-like protein 1,growth hormone d6,mature ? chain of Mhc-bi protein,A-1 inhibitor 3,complement C1 s and other five up-regulated proteins,glutathione.Peptide S-transferase,Ube2l3 protein,creatine kinase type B,ubiquitin-binding enzyme e2 v1,glucose-6-phosphate1-dehydrogenase,cysteine and glycine-rich protein 1,ADP-ribosylation factor 3.Peptidyl proline sequence 9 enzymes such as FKBPIA and 14-3-3 protein ? aredown-regulated.3.Bioinformatics analysis: 1 From the GO function annotation analysis,it is found that the biological processes involved in differentially expressed proteins include metabolic processes,cellular processes,biological regulation,and responses to stimuli.It has molecular functions such as binding and catalytic activity.In the cell components,mainly in the cell part,the extracellular part,the organelle and the organelle part.From GO functional enrichment analysis(P<0.05,TOP10):positive regulation of endocytosis from biological processes,sputum development,positive regulation of I-Kappa b kinase/NF-kappa B signaling,response to food,etc.Molecular function is mainly enriched in ubiquitin-binding enzyme activity,ubiquitin-protein ligase activity,etc.,and is enriched in ubiquitin ligase complexes on cellular components.2 From the KEGG pathway annotation and enrichment analysis,we found 51 signal pathways,of which P<0.05 pathway has glutathione metabolism,endocytosis,prolactin signaling pathway,Toll and IDM signaling pathway,virus carcinogenesis,etc.5 channels.Signal transduction pathways such as glutathione metabolism,prolactin signaling pathways,and viral carcinogenesis are associated with anti-aging in these five pathways.Conclusion: 1.Mongolian medicine Zhenhe moxibustion therapy achieves the goal of anti-aging by influencing the related enzymes,physical strength,learning and memory of model rats.2.Fourteen differentially expressed proteins,including fibrinogen-like protein 1,growth hormone d6,mature alpha chain of Mhc-ib proteins,AI-1-inhibitor 3 and complement C1 s,were screened.Bioinformatics analysis of differentially expressed proteins was carried out to understand biological processes,molecular functions and the relationship between existing cellular components and their differentially expressed proteins.The related signaling pathways are used to fight aging.3.This study further proves the principle of anti-aging acupoint allocation of Mongolian medicine Zhenhe moxibustion,and also achieves the purpose of explaining the clinical mechanism of anti-aging of Mongolian medicine moxibustion with protein and its pathway,and provides a scientific basis.