| ObjectiveThe paper explores the associated proteins of the hypothalamus. in aging rats which induced by D-galactose with intervention by Qiongyugao, and find out the target protein candidates by bioinformatics analysis. Hence we can research the mechanism of the action of Qiongyugao at the molecular leve, and provide a theoretical basis for the development of Qiongyugao.MethodsThe 42 6-month-old male SD rats are divided randomly into three groups(Qiongyugao group, control group, model group), each group has 14 rats with 2 cages. From the first day of the experiment, the model group were injected intraperitoneally with D-galactose(0.3g/kg) every day in order to manufact subacute aging model, and the same Qiongyugao with model group, the control group is traeted with saline at the same way with the other two groups;Meanwhile the rats of Qiongyugao group are fed into stomach with Qiongyugao 0.583ml/100g every day, and the ones of model and control groups are fed saline at the same way, the experimental process is continued for 40 days. After 40 days, the serum and liver in the three groups’rats is got for antioxidant capacity tests.At the same time, full protein in hypothalamus is got for iTRAQ study, with the process of protein enzymolysis, labeled peptides, high pH reverse phase chromatography, mass spectrometry and other methods to determine significant differences in protein. Combining with Gene Ontology annotations, KEGG and protein networks related bioinformatic analysis, the paper studys the function, properties,metabolic pathways and network of the differences in protein, and find out the target protein candidates.ResultsWe made model of aging rats successfully, and find out that Qiongyugao can increase the activity of SOD in rat’s liver and GSH-Px in serum, the result is of significance in statistics science.Through analysis of proteomics technology, there are 14 kinds of significant differences in proteins which are identified as Cytochrome c oxidase polypeptide VIc-2, Neutral ceramidase, Phytanoyl-CoA hydroxylase-interacting protein-like, Nicotinanamide phosphoribosyl transferase, Tail-anchored protein insertion receptor WRB, Protein O-linked-mannose beta-1,4-N-acetylglucosaminyl transferase 2, Signal transducer and activator of tran-scription 5B,Acetoacetyl-CoA synthetase, Tubulin-specific chaperone E,3-phosphoinositide-dependent protein kinase 1,Dual specificity mitogen-activated protein kinase kinase 7, RAS guanyl-releasing protein 2, SHC-transforming protein 3, Pantothenate kinase 4, and Qiongyugao play the role of these proteins are down-regulated.Through analysis of bioinformatics analysis, there are 31 kinds of functions associated with these differences in proteins, including 15 kinds of biological processes,9 kinds of cells and molecular function 7 kinds. KEGG pathway analysis reveales that there are 5 major metabolite pathways, respectively is Fc e RI signaling pathway, Pantothenate and CoA biosynthesis, T cell receptor signaling pathway, ErbB signaling pathway and Sphingolipid metabolism.ConclusionThe 14 kinds of significant difference proteins includes mitochondrial proteins, metabolism related protein, growth realted protein, the protein involving in connection, catalytic related proteins and signal transduction related proteins. Through bioinformatics analysis, the target proteins in which the Qiongyugao infuences the hydroxylase of aging rat are some of the 6 kinds of proteins which are Neutral ceramidase, Phytanoyl-CoA hydroxylase-interacting protein-like, RAS guanyl-releasing protein 2, SHC-transforming protein 3, Dual specificity mitogen-activated protein kinase kinase 7, Signal transducer and activator of transcription 5B, corresponding genes are Asah2, Phyhipl, Rasgrp2, Shc3, Map2k7 and Stat5b. |
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