Differences Expression Proteins Of HIV/AIDS Deficiency Syndrome Of Lung-Spleen Qi Based On ITRAQ-MS Technique

Objective: Searching for differentially expressed proteins of HIV/AIDS deficiency syndrome of lung-spleen qi use proteomics technology. By means of bioinformatics analysis of differentially expressed proteins, studying the relationship between it and clinical syndrome of HIV/AIDS deficiency syndrome of lung-spleen qi, exploring the biological basis of HIV/AIDS deficiency syndrome of lung-spleen qi.Methods: 1 25 cases of HIV/AIDS deficiency syndrome of lung-spleen qi patients were collected by the diagnosis scale of HIV/AIDS deficiency syndrome of lung-spleen qi,16 cases of healthy people with the same area as control group, collect the serum samples of the research object; 2 Use multiple affinity centrifugal removal system Human-14 to get rid of 14 kinds of high abundance proteins, then use SDS-PAGE gel electrophoresis to evaluation for removal efficiency of high abundance proteins; 3 Proteins in serum were analyzed using i TRAQ-MS technique, and the mass spectrometry data were appraisal analysis by Mascot software to serch uniprot. Human. Fasta database, and peptides report ion peak intensity values were quantitative analysis use the Proteome Discoverer software, then screen differentially expressed proteins of HIV/AIDS deficiency syndrome of lung-spleen qi; 4 Bioinformatics analysis using NCBI BLAST + localization sequence alignment software to identify proteins and compare the human protein sequences in the Swiss Prot database, use of the Mapping Function of Blast2 GO to progressive target protein sequence alignment with the associated gene ontology(GO) functional item extraction, analysis of the target protein involved in the Biological processes(BP), Molecular Function(MF), cell components(CC).Using KAAS(KEGG Automatic Annotation Server) compare target protein sequence with the sequence of human proteins in the KEGG GENES database, by the same/similar protein of KO(KEGG Orthology) comments on the KEGG pathway, analyze which metabolism and signal transduction pathways the target proteins were involved.Results: 1 There is no obvious difference of age, sex composition and CD8 levels between the two groups(p>0.05),but of HIV/AIDS deficiency syndrome of lung-spleen qi patients CD4, CD4 / CD8 levels were significance in low stage than control group(p<0.05); 2 SDS-PAGE gel electrophoresis results shows after removal of high abundance protein of serum the electrophoresis banding increased remarkably,can significantly reduce the bridging effect of high abundance proteins on the low abundance, also different molecular weight protein separation clear; 3 After removal of high abundance protein and enzyme digestion, i TRAQ reagent tag, LC-MS analysis, Mascot libraries were identified 2612 Unique Peptide, 279 Protein Group, including 195 proteins with quantitative information of all channels; 4 Ratio change between group in mass spectrometry results together with t-test analysis P value for screening differentially expressed proteins:1. To ratio changes> 1.2 or < 0.83,p<0.05 for the standard, 4 proteins were identified to be differentially expressed, 2 were up regulation expression, 2 were down regulation expression;2. With ratio change> 1.15 or < 0.87,p<0.05 for the standard, 8 proteins were identified to be differentially expressed, 3 were up regulation expression, 5 were down regulation expression; 5 Only with the standard p values< 0.05 that 24 protein were to be differentially expressed,bioinformatics analysis showed that they involve biological process include killing, biological cell adhesion, multicellular organization process, biological process single organ, cell migration, replication total nine aspects;Molecular functions include receptor activation, transshipment activation, combining total three aspects;Cell component include membrane, cell, extracelluar region, membrane-enclosed lumen, macromolecular mixture total five aspects; 6 KEGG pathway analysis showed that differentially expressed proteins mainly involve 10 metabolism or signaling pathways including: systemic lupus erythematosus(sle), Fc-gamma R mediated phagocytosis, infection of staphylococcus aureus, fat digestion and absorption, vitamin digestion and absorption, receptor activation and viral carcinogenesis, prion disease, complement cascade coagulation, amoebiasis.Conclusion: 1 Multiple affinity centrifugal removal system Human-14 can effectively remove the 14 kinds of high abundance proteins in serum, reduce the bridging effect of high abundance proteins on the low abundance proteins, that can be a advantageous to the discovery of biomarkers associated with disease, and helpful for the study of the biological basis of syndromes; 2 i TRAQ-MS technology could screen for differentially expressed proteins of HIV/AIDS deficiency syndrome of lung-spleen qi, prompting it has a good prospects in the use of HIV/AIDS related biomarkers of different syndromes.Differentially expressed proteins in the occurreence and development of the disease has not yet entirely clear, further study should be done to evaluate its reliability 3 Chinese syndromes as a specific stages of the evolution of the disease symptoms and signs of generalization has biological basis.Bioinformatics analysis of differentially expressed proteins revealed that the differentially expressed proteins function and signal, metabolic pathways associated with immune and nutrient digestion and absorption.The abnormal express of urea transporter, apolipoprotein A-II and zinc – alpha-2-glycoprotein are closely related to the symptoms of HIV/AIDS deficiency syndrome of lung-spleen qi: cough phlegm thin, weak asthma, bosom frowsty, flustered, heart palpitations, diarrhea and so on,the three proteins are likely to be potential biological basis of HIV/AIDS deficiency syndrome of lung-spleen qi 4 Zinc-alpha-2-glycoprotein this study identify differentially expressed proteins, one of its protein and gene expression in malignant tumor of lower level involves multiple organs, and lipid metabolism disorder, the literature only reports that zinc-alpha-2-glycoprotein as one kind of tumor biological markers is one-sided. Zinc-alpha-2-glycoprotein expression can explain why the patients with HIV/AIDS deficiency syndrome of lung-spleen qi have high blood fat in circulation system,can also predict the possibility of later developed into the tumor, both in the theory of Chinese medicine and in modern pathology study can be found that zinc-alpha-2-glycoprotein is likely to be HIV/AIDS deficiency syndrome of lung-spleen qi a series of biological basis of pathology of evolution.