Differentiation Analysis Of Serum Proteins In Rat Under The Subchronic Smoke Exposure

Objectives:In recent years, the incidence and mortality caused by lung related diseases are increasing year by year. This thesis was focusing on the proteomics analysis, how cigarette or air pollution influence the proteins of lung tissues exposed cicigarette smoke on rat model.Methods:160 rats were used to establish animal model and divided into control groups and 3 treatment groups with three different cigarette smoke concentrations. The smoke concentration was set as 10%,20%, and 60%, and the time was 3 and 6 months. The lung tissues were separated and treated by serials of protein separation processes. Then the protein was digested by enzymes and followed by using theiTRAQ to identify and analyze the peptides structure and components. Proteins from samples of four groups (CM/LM/MM/HM) was initially digested by FASP, iTRAQ labeling, RPLC separation, and finally analyzed by RPLC-MS. The data collected was further matched with database.Results:The rat model was successfully established, and the data of 340258 ions,19094 peptides and 4483 proteins was collected. Statistical distribution of peptides was analyzed according to the molecular weight. The quantitative analysis showed a diverse expression among the four groups. Compared to control group, the proteins from the other three groups showed a tendency of dose-dependent according to our annotation results. GO Annotation suggested that the functional proteins comprise three aspects:mechanism pathway, cell adhesion, cell processing. COG annotation showed that the identified proteins can be concentrated into 26 sub-groups. KEGG database analysis demonstrated that there have different number of signal-pathway related proteins in the three groups with different doses of smoke compared with control.Conclusions:iTRAQ combined mass spectrometry was used to analyze lung tissue proteins to exafter cigarette smoke exposure on Wistar rats. Our results suggested there ware significant changes on proteins after the subchronic smoke exposure which showed dose-dependent. Those proteins identified showed that themetabolism, PPAR pathway mechanism, Hypertrophic cardiomyopathy, dilated cardiomyopathy, and myocardial infarction were involved. Our preliminary results provide some new insights of detecting and diagnosis for lung diseases, and some identified proteins with significant differences can also be candidate for biomarkers which need further investigation.