Circ_PWWP2A promotes lung fibroblast proliferation and fibrosis via the miR-27b-3p/GATA3 axis, thereby aggravating idiopathic pulmonary fibrosis.

Abstract

Objective: This paper was to investigate the effect of circ_PWWP2A-mediated miR-27b-3p/GATA3 axis on idiopathic pulmonary fibrosis (IPF).

Methods: circ_PWWP2A expression in lung fibroblasts MLg2908 induced by different concentrations of TGF-β was detected. The relationship between circ_PWWP2A or GATA3 and miR-27b-3p was analyzed by RNA immunoprecipitation and dual-luciferin reporter assay. The proliferation of MLg2908 cells was determined by MTT. GATA3, α-SMA, Collagen-I, and Collagen-III in cells were detected by RT-qPCR and Western blot. The rat model of IPF induced by bleomycin (BLM) was constructed and treated with circ_PWWP2A siRNA injection. HE and Masson staining were of utility to evaluate the pathological conditions of rat lung tissue, and circ_PWWP2A, miR-27b-3p, and GATA3 levels in lung tissues were detected by RT-qPCR. Immunohistochemistry was used to detect the staining of α-SMA, collagen I, and collagen III in the lung tissues of rats.

Results: circ_PWWP2A in MLg2908 cells induced by TGF-β decreased in a concentration-dependent manner. MLg2908 cells transfected with circ_PWWP2A siRNA were induced by 5 ng/ml TGF-β, decreasing circ_PWWP2A and GATA3 levels, increasing miR-27b-3p expression, and suppressing cell proliferation. The targeting relationship between circ_PWWP2A and miR-27b-3p, as well as miR-27b-3p and GATA3, was confirmed. Depleting miR-27b-3p reduced the inhibitory effect of circ_PWWP2A down-regulation on the proliferation of TGF-β-treated MLg2908 cells, accompanied by increased expression of α-SMA, Collagen 1, and Collagen 3, and increased expression of GATA3. The in vivo results showed that BLM-induced fibrosis in rat lung tissue was obvious, accompanied by increased expression of circ_PWWP2A and GATA3, decreased expression of miR-27b-3p, and deepened staining of α-SMA, collagen I, and collagen III, but circ_PWWP2A siRNA could improve these phenomena.

Conclusion: Silencing circ_PWWP2A can inhibit the proliferation of lung fibroblasts induced by TGF-β through the miR-27b-3p/GATA3 axis, and reduce BLM-induced pulmonary fibrosis in rats, which may be a potential therapeutic target for IPF.