Puerarin mediated miR-30b-5p targeting fibroblast activation protein against oral submucous fibrosis

Background: Puerarin (Pue) has been reported to be a natural active ingredient with multiple antifibrotic properties. This work aimed at exploring the function of Pue in oral submucous fibrosis (OSF) treatment. Methods: Human oral mucosa fibroblasts (hOMF) were induced with transforming growth factor beta1 (TGF-(31) and intervened with Pue. Expressions of fibrosis-related markers were analyzed by Western blot and IF staining. Cell viability was characterized by the CCK-8 assay. Expressions of miR-30 family members were quantified by qRT-PCR. The correlation between fibroblast activation protein (FAP) and miR-30 family expression was evaluated by the Pearson correlation coefficient. Bioinformatics prediction and dual-luciferase reporter assay were employed to verify the regulation between FAP and miR-30b-5p. The specific mechanism of Pue on OSF was explored through the promotion or inhibition of miR-30b-5p. Results: After induction by TGF-(31, hOMF showed upregulated Collagen I, Collagen III, and FAP expressions, while miR-30 family expression was downregulated with miR-30b-5p being the most significant. Pue intervention inhibited the excessive proliferation of TGF-(31-induced hOMF, downregulated FAP, collagen type 3 (COL3A1), collagen type 1 (COL1A1), matrix metalloproteinase 1 (MMP1), and matrix metalloproteinase 3 (MMP3) expressions, and restored miR-30 family expression. Bioinformatics prediction and dualluciferase reporter assay revealed that miR-30b-5p selectively inhibited FAP expression. Mechanistically, miR-30b-5p mimic suppressed the excessive proliferation of TGF-(31-induced hOMF and declined fibrosis levels. Pue intervention significantly reversed the promotion of TGF-(31-induced OSF by miR-30b-5p inhibition. Conclusion: Pue mediated miR-30b-5p targeting FAP against OSF, which provided a theoretical basis for the pathogenesis research and Pue application in OSF.