The Mechanism Of DsbA-L Up-regulating The Expression Of AKT1 And NLRP3 To Induce Pulmonary Fibrosis

Objective: Up to now,the pathogenesis of pulmonary fibrosis is not fully understood.The damage of alveolar epithelial cells plays a crucial role in the occurrence of pulmonary fibrosis,and mitochondrial damage is a key factor in the occurrence of alveolar epithelial damage.Disulfide bond A oxidoreductase-like protein(DsbA-L)is highly correlated with cellular oxidative stress and mitochondrial damage.This study aimed to explore whether DsbA-L plays a role in pulmonary fibrosis and to explore its related molecular mechanism.Materials and Methods: Firstly,the expression difference of DsbA-L in pulmonary fibrosis tissue and normal tissue was analyzed by GSE24206 dataset in GEO database.Western blot experiments were performed to explore the differences in the expression of DsbA-L in human pulmonary fibrosis tissue samples and normal lung tissue samples.Then,TGF-β1 was used to intervene alveolar epithelial cells,and the expression of DsbA-L was detected.A cell model overexpressing DsbA-L and silencing DsbA-L was constructed by transfecting plasmids into alveolar epithelial cells,and TGF-β1 was used to intervene the cell line.Expression of DsbA-L,AKT1,Smad3 and phosphorylated Smad3.Finally,the regulatory relationship between TGF-β1,DsbA-L,AKT1/NLRP3 was further clarified by transfecting alveolar epithelial cells with AKT1 and NLRP3 agonists.Results:(1)Analysis of the GSE24206 dataset showed that DsbA-L expression was increased in fibrotic lung tissue compared to normal lung tissue.(2)The detection results of human tissue samples showed that the expression level of DsbA-L in fibrotic lung tissue was higher than that in normal lung tissue.(3)After adding TGF-β1 to intervene alveolar epithelial cells,the expression level of DsbA-L increased with the prolongation of intervention time.(4)The expression level of DsbA-L was significantly increased in the cell lines transfected with overexpressing DsbA-L plasmid,while the expression level of the silencing group decreased.After the intervention of TGF-β1,the levels of AKT1 and NLRP3 in DsbA-Loverexpressing cell lines were significantly increased,and the expression levels of type I and type III collagen fibers were increased.However,the expression levels of AKT1 and NLRP3 in the cell lines that silenced DsbAL decreased,and the expression levels of type I and type III collagen fibers were reduced.(5)After adding AKT1 and NLRP3 agonists,the expressions of phosphorylated Smad3 and TGF-β1 were further increased.Conclusions:(1)The expression level of DsbA-L in fibrotic lung tissue is higher than that in normal lung tissue;(2)TGF-β1 induces the expression of DsbA-L in alveolar epithelial cells;(3)DsbA-L up-regulates the expressions of AKT1 and NLRP3,and DsbA-L might mediates the TGF-β/Smad3 signaling pathway to promote pulmonary fibrosis.Figures 12 Tables 4 References 71...