Differential Protein Study Of Alveolar Macrophages In Different Stage Silicosis Patients

Objectives To obtain protein related with the progress of occurrence and development of silicosis and further reveal the development mechanism of silicosis,we collected alveolar macrophages in the lung lavage fluid of silicosis patients and investigated the different protein in alveolar macrophages from different stage silicosis patients.Methods The subjects were selected from silicosis patients,in China's coal miners Beidaihe sanatorium located on Beidaihe from October 2016 to September 2017.We choosed workers exposed only to rock dust as research subjects.The subjects suffered from as few other diseases as possible.At present,they were still Finally,we selected 31 cases of silicosis who are male and exposed to dust,or dust removal within 2 years.The subjects were composed of 5 cases of pulmonary dust deposition,10 cases of silicosis on grade one,two cases of silicosis on grade two,9 cases of silicosis on grade three,and 7 cases of silicosis grade three.After consulting the records of the medical records,we selected the subjects that met the conditions of admission and signed the informed consent book,and collected the first 8 bottles of lavage fluid in the patients with silicosis and extracted the macrophages in the lung lavage fluid.The macrophages from first 8 bottles of lung lavage fluid were divided into two parts.The macrophages from first 7 bottles of lung lavage fluid were used for extraction of total protein on ice.The proteins were stored up in-80? refrigerator.The macrophages from the eighth bottle of lung lavage fluid were used for Switzerland's Jim dyeing for morphologic observation.Two dimensional gel electrophoresis was used to isolate the differential proteins.Image Master 2D Platinum software was used to identified differential protein spots.This method was used to identify differential spots,namely New ultrafle Xtreme reflective tandem matrix assisted laser desorption time of flight mass spectrometry.GO,KEGG Pathway and PPI analysis of differential proteins were carried out by bioinformatics.Results The ten factors,age,area,smoking history,drinking history,dust exposure time,dust property,COPD,complicated by fatty liver and nephrolithiasis,and cardiovascular and cerebrovascular diseases,were evenly distributed among different stages of silicosis and were comparable.After analyzed by Image Master 2D Platinum software,the difference between the first grade silicosis group and the lung dust deposition group was the proteins which Match ID was 3033,2789,2726,2649,3207,3008.The difference between the second grade silicosis group and the lung dust deposition group was the proteins which Match ID were 3516,3304,2276,1792,3008,1821,2967,2805,2376,1857,1349.The difference between the third grade silicosis group and the lung dust deposition group was the proteins which Match ID were 2828,2863,3681,3704,2959,2965.By New type ultrafle Xtreme reflection type series matrix assisted laser desorption time of flight mass spectrometry,we identified seven kinds of protein,which Match ID were 2789,3033,2967,3516,3304,3008,1857,respectively.The seven kinds of protein,were ANXA4,PRDX4,ERP29,KRT10,FTL,ECHS1,ALDH2.The above results enrich the differential proteomics database of silicosis patients.Through bioinformatics analysis,the main functions of these proteins include protein connection,activation of aldehyde dehydrogenase,binding of calcium dependent proteins,activation of phospholipase inhibitors,and binding of iron ions,etc.The main biological processes of these proteins include fatty acid beta oxidation,alcohol metabolism,regulatory gene expression and secretion,activation of MAPK activity,active oxygen metabolism,neutrophil degranulation,Notch signaling pathway,and negative regulation of interleukin-8 secretion,etc.These proteins are the main components of the self phagocytic bubble,the matrix of the mitochondria,the cell membrane,the cytoplasm,the endoplasmic reticulum,the transport vesicles,the inner cavity of the secretory granules,the exocrine and so on.Through bioinformatics analysis,these differential proteins mainly participate in the pathway of fatty acid degradation,valine,leucine and isoleucine degradation,lysine degradation,tryptophan metabolism,pyruvate metabolism,metabolic pathways,and so on.Bioinformatics analysis showed that there were interactions between ECHS1 and ALDH2.Conclusions 1 Using differential proteomics,we found that the proteins associated with silicosis development in alveolar macrophages were ANXA4,PRDX4,ERP29,KRT10,FTL,ECHS1,ALDH2,which enriched the alveolar macrophage protein library of silicosis.2 Through bioinformatics analysis,the main mechanisms of pulmonary alveolar macrophages involved in the development of silicosis were fatty acid metabolism,MAPK activity regulation,oxidative stress,notch signaling pathway,and negative regulation of interleukin-8 secretion.3 The expression of ECHS1 protein in the pulmonary dust deposition group was different from the silicosis groups of grade one and two.ECHS1 protein may be the key protein in the development of silicosis.