Exploring The Potential Effective Signaling Pathway Of Silicosis Based On Bioinformatics Analysis And Verifying The Function Of C-Jun In Silica-induced Inflammation

Objective: Silicosis is a pneumoconiosis caused by long-term inhalation of free silicon dioxide dust,with diffuse fibrosis in the lung,which the fibrosis lesions damage the lung function,and finally leading to respiratory failure.It is still the focus of silicosis research to study the pathogenesis of silicosis and find effective treatment methods.The purpose of this study is to deeply analyze the single cell sequencing data of silicosis patients based on bioinformatics analysis,explore the potential pathogenesis of silicosis and to verify it by experiments.In order to provide effective basis for the treatment of silicosis.Methods: In this study,the GSE174725 data set was selected and integrated by R language programming software.the appropriate sample data of silicosis patients(case data group)and control sample data(control data group)were selected and integrated into new dataset.Using Sub Set grammar of R for quality control.Using UMAP function to perform cell clustering analysis.Cell Chat function was used for cellular communication analysis.Analyzing the protein-protein interaction network by string database and defining the Hub gene in the network.Online prediction of the downstream target genes of the Hub gene by JSPAR database,and the KEGG signaling pathway and GO function of the Hub gene is clarified in combination with the Differentially expressed genes.HE and Masson staining techniques were used to clarify the pulmonary inflammation and fibrosis in the case group and the control group.Immunohistochemical staining technique was used to explore the expression of c-Jun in lung tissue of case group and control group.THP-1 cell line was used for experimental verification in vitro.Silica stimulated THP-1 cells for 24 hours after 1-hour pretreatment with cJun specific inhibitor SR11302.Realtime-PCR was used to detect the m RNA expression of c-Jun,TNF-α,IL-1β and IL-6.Results:1.The results of data quality control show that after quality control,the gene content of each cell is less than 2500,the mitochondrial gene is 0,and the ribosome gene is more than 2.5%.quality control was successed.2.Cellular analysis showed that the proportion of macrophages in the case group was significantly higher than that in the control group,and the proportion of monocytes was significantly lower than that in the control group.MIF mediated signaling pathway plays an important role in cellular communication.3.Protein-protein interaction network analysis showed that c-Jun is one of the core protein of the regulatory network in silicosis,and c-Jun could regulate the TLRs signaling pathway and bind the inflammatory receptor proteins in silicosis.4.C-Jun is highly expressed in lung tissue of silicosis patients,and specific inhibition of c-Jun can reduce the expression of TNF-α and IL-6 m RNA in THP-1 cells caused by silica in vitro.Conclusion:1.MIF mediated signaling pathway may affect immune cell interaction in silicosis patients through D74/CD44 and CD74/CXCR4ligand-receptor.2.C-Jun transcription factor may activate TLRs signaling pathway by inducing downstream target genes during the development of silicosis.3.Inhibition of c-Jun can alleviate the inflammatory reaction of THP-1 cells induced by silica in vitro.