Research On Non-coding RNA Regulatory Networks In The Development Of Pulmonary Fibrosis

Objective:Pulmonary fibrosis is a special form of chronic,progressive fibrosis of interstitial pneumonia.The mechanism is unknown and there is no cure.Noncoding RNAs(ncRNAs),including miRNA,lncRNA,and circRNA,they are regulators of important biological functions.By studying the regulation of ncRNA,finding a gene therapy target for the prevention and treatment of pulmonary fibrosis or a biomarker for the diagnosis and treatment of disease,and provide a theoretical basis and development of corresponding intervention drugs in the future,so that the disease can be Early detection,early diagnosis,early treatment,improve the quality of life of patients,improve survival rate,and reduce the burden on clinical medical and nursing staff.Methods:Differentially expressed RNA in bleomycin-induced animal model of pulmonary fibrosis was screened using whole transcriptome sequencing.Four differentially expressed miRNAs,ten lncRNAs and two circRNAs were detected in the cells by qRT-PCR assay to verify RNA sequencing results.Characterization of differentially expressed ncRNAs and mRNAs and construction of co-expression networks by bioinformatics techniques.The regulation of ncRNAs on their host genes and target genes on pulmonary fibrosis was studied by using RNA fluorescence in situ hybridization(RNA-FISH),dual luciferase reporter assays,Western Blot and Real Time Cellular Analysis(RTCA).Results:Differentially expressed 585 mRNAs,236 miRNAs,272 lncRNAs and 74 circRNAs were screened.Lnc949,circ949 and circ057 were selected as subjects for exploring the regulation pattern of ncRNA in pulmonary fibrosis.RNA-FISH showed that all three ncRNAs predominate in the cytoplasm,and their regulatory patterns are mainly concentrated at the post-transcriptional level.The profibrotic function of lnc949 is mainly determined by its host gene FKBP5.Circ949/circ057 and lnc865/lnc556 form an RNA regulatory network targeting miR-29b-2-5p to jointly regulate the STAT3 gene in pulmonary fibrosis.Conclusion:Different RNAs can crosstalk with each other to regulate pulmonary fibrosis through different regulatory patterns.