| Objectives To obtain the main protein types and quantities of silicosis and its complications,and to analyze the protein function related to silicosis by comparing the differences in protein expression in bronchoalveolar lavage fluid between patients with silicosis in different periods and controls,combined with Bioinformatics analysis,Signal pathways and interactions provide ideas for understanding of the pathogenesis of silicosis.Method There were 38 workers who had previously undergone large-volume lavage treatment and had not received large-volume whole-pulmonary lavage treatment in the mining industry,including 19 cases in period(40),5 cases of period ? and 14 cases of period ?.The control group There were 5 patients with negative bronchoscopy results in control group and without hepatitis B or other pulmonary fibrosis.With the informed consent of the person or family,the second bottle of lung lavage fluid in silicosis group and the lavage fluid of the normal lung tissue of the control group were collected and frozen at-80 °C.Pretreat lung lavage fluid by 10 K ultrafiltration tube ultrafiltration,Agilent 1100 conventional liquid phase separation,strong cation exchange chromatography column(SCX)HPLC pre-separation and C18 reverse phase chromatography desalination purification,using isotope-labeled protein.Mass spectrometry was performed on a Thermo Q-Exactive mass spectrometer.The mass spectrograms were screened using Proteome Discoverer 1.4 thermo software,searched using mascot,and protein differential analysis was performed based on mascot search results and post-screen spectra to determine differential proteins.Apply GO analysis,KEGG Pathway analysis,and PPI analysis of different proteins to obtain corresponding signaling pathways and interaction.Results The age,the history of smoking,the time of picking up dust,nature of the dust,COPD,total five factors from selected 43 cases were uniform distribution in different stages of silicosis.943 kinds of proteins were identified by Thermo Q-Exactive Mass Spectrometry.A total of 226 differential proteins were obtained between the period(40)groups and control groups,102 kinds of proteins were up-regulated and 124 kinds of proteins were down-regulated.A total of 234 differential proteins were obtained between the period ? groups and control groups,118 kinds of proteins were up-regulated and 116 kinds of proteins were down-regulated.A total of 205 differential proteins were obtained between the period ? groups and control groups,97 kinds of proteins were upregulated and 108 kinds of proteins were down-regulated.A total of 229 differential proteins were obtained between the period ? groups and period ?groups,107 kinds of proteins were up-regulated and 122 kinds of proteins were down-regulated.A total of 138 differential proteins were obtained between the period ? groups and period(40)groups,70 kinds of proteins were up-regulated and 68 kinds of proteins were down-regulated.A total of 207 differential proteins were obtained between the period ? groups and period(40)groups,111 kinds of proteins were up-regulated and 96 kinds of proteins were downregulated.differential proteins includes Zymogen granule protein 16 homolog(BA0A0C4DGN4),Serum amyloid P-component(P02743),Calreticulin,(B4DHR1),Apolipoprotein A-II(P02652),Epididymis secretory protein Li 55,(V9HW35),Calcyphosin(Q13938),Peroxiredoxin-6(P30041),Antileukoproteinase(P03973).By bioinformatics,the main pathways of differences protein spots were Metabolic pathway,Proteasome,Glycolysis/Gluconeogenesis,Viral carcinogenesis,Systemic lupus erythematosus,Estrogen signaling pathway,Endocytosis,Chemical carcinogenesis,Epstein-Barr virus infection,PI3K-Akt signaling pathway,Pyruvate metabolism,IL-17 signaling pathway,HIF-1 signaling pathway,PPAR signaling pathway.Conclusions 1 The proteomics method using isotope relative and absolute quantification(iTRAQ)combined with Q-Exactive mass spectrometry found that there were 943 proteins related to the development of silicosis in bronchoalveolar lavage fluid of patients with silicosis in different stages,a total of 491 species.2 The main biological changes in the early stage of silicosis are glycolysis/gluconeogenesis,autoimmunity,carbon metabolism,phagocytosis,etc.and microfibrillar-associated glycoprotein 4 may be involved in the early biological processes of silicosis.3 The main biological changes in the late stage of silicosis are autoimmune,intercellular adhesion,and calcium hippocampal binding protein may be involved in the biological process of late silicosis.Figure16;Table16;Reference 142... |
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