CircRNA-102348 Protects Target TAZ In Idiopathic Pulmonary Fibrosis By Adsorbing MiR-630

Objective:Idiopathic pulmonary fibrosis(IPF)is a typical senile disease,usually with a poor prognosis and no effective treatment.It is characterized by clinical radiology and pathophysiology models,and usually has a poor prognosis and no effective treatment drugs.Through a new perspective to explore the pathogenesis of IPF,the discovery of new non-invasive biological markers and gene drug treatment targets with early diagnostic value is an important research area of respiratory diseases,which has important social and economic value.The purpose of this paper is to clarify the mechanism by which circRNA-102348 protects the target gene TAZ in idiopathic pulmonary fibrosis by adsorbing miR-630,and to reveal the way of transmission of genetic information mediated by non-coding genes and the regulatory model of circRNA-protein complexes provides targets for gene therapy and drug design of IPF.Methods:The tranRNAs differentially expressed in the blood of IPF patients and normal humans were screened by full transcriptome sequencing technology.Using Sanger sequencing to determine the circularization site of circRNA-102348.Design interference fragments and construct overexpression vectors based on full-length sequences.TGF-β1-induced differentiation of human fibroblast cell line MRC-5 into lung fibrosis cell model;siRNA and overexpression vectors were used to knock down and increase circRNA-102348,respectively.Using Western Blot and Real-Time Cell Analysis System(RTCA)to detect changes in myofibroblast activation and function-related indicators under the intervention of circRNA-102348;Using RNA-FISH to detect the intracellular localization of circRNA-102348 and determine the cell’s regulatory pattern;Ago RIP and dual luciferase reporter gene experiments were used to further determine their binding relationship;The effect of miR-630 on activation,proliferation,migration,and collagen secretion of myofibroblasts was measured by Gain/Loss of function.Predict and verify the targeting relationship between miR-630 and TAZ through mass spectrometry analysis,database analysis,and double luciferase reporter gene experiments;Rescue experiments verified the protective effect of circRNA-102348 on TAZ by miR-630 adsorption.Results:The results of whole transcriptome sequencing technology showed that circRNA-102348 was significantly overexpressed in the blood samples of IPF patients,and the expression level of circRNA-102348 had significant clinical correlation with IPF patients;circRNA-102348 can promote the activation,proliferation,migration,and collagen secretion of myofibroblasts;circRNA-102348 has a targeting relationship with miR-630,miR-630 and TAZ.Rescue experiments have demonstrated the protective effect of circRNA-102348 on TAZ,and circRNA-102348 forms an RNA regulatory network targeting miR-630 to protect target gene TAZ.Conclusion:circRNA-102348 has significant correlation with IPF,and circRNA-102348 can protect the target gene TAZ by adsorbing miR-630.The presence of circRNA-102348/miR-630/TAZ axis as a gene delivery system is involved in regulating the occurrence and development of IPF.