Mechanism of miR-15a-5p regulation of Wnt pathway in paraquat-induced pulmonary fibrosis
Objective To investigate the effect and molecular mechanism of miR-15a-5p regulation Wnt signaling pathway in PQ-induced pulmonary fibrosis.Methods The PQ-induced 16HBE cell model was constructed,high-throughput miRNA chip and RT-qPCR were used to screen for miR-15a-5p with significant differences.The experimental groups were as follows:NC group(normal control);no special treatment;PQ group:50 μmol/L PQ treated cells for 72 h;miR-15a-5p group:16HBE stable cell lines transfected with miR-15a-5p overexpressing lentivirus;miR-15a-5p+PQ group:Stable cell lines were treated with 50 μmol/L PQ for 72 h.The expression of Wnt pathway-related genes Wnt3α andβ-catenin,fibroblast marker genes Collagen I,Vimentin and α SMA,epithelial marker genes Occludin and CK18 were detected by RT-qPCR and Western blot.The mice model of PQ-induced pulmonary fibrosis was constructed,and the protein expression and lung tissue injury were detected by Western blot,HE staining and immunohistochemistry.Data were expressed as mean±standard deviation,and independent sample t-test was used to analyze the data between the two groups.Results The expressions of Wnt3α,β-catenin,fibroblast marker genes Collagen I,Vimentin and α SMA significantly up-regulated in cell injury models(P＜0.05),the epithelial cell marker genes Occludin and CK18 significantly down-regulated(P＜0.05),overexpression of miR-15a-5p could inhibit the expression of Wnt3α and alleviated the EMT induced by PQ.In animal models,Wnt3α,β-catenin,fibroblast marker genes Collagen I,Vimentin and αSMA significantly increased(P＜0.01),the structure of lung tissue was disordered and fibrosis occurred,overexpression of miR-15a-5p inhibited the expression of Wnt3α protein(P＜0.05)and ameliorated lung tissue injury.Conclusions miR-15a-5p ameliorates PQ-induced lung injury by modulating the Wnt3α/β-catenin signaling pathway,thereby inhibiting the development of pulmonary fibrosis.
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