Tanshinone IIA down-regulated p-Smad3 signaling to inhibit TGF-β1-mediated fibroblast proliferation via lncRNA-HSRL/SNX9

Abstract

Introduction: Tanshinone IIA (TSIIA), an active component of Salvia miltiorrhiza (Danshen), is reported to inhibit cell proliferation in hypertrophic scars (HS). In our previous study, we observed that lncRNA human-specific regulatory loci (HSRL) was up-regulated in HS tissues. However, whether TSIIA serves as an effective treatment for HS through affecting HSRL is still unexplored.

Methods: TGF-β1-stimulated fibroblast were used as the in vitro HS model. The effects of TSIIA on cell proliferation were evaluated using CCK-8, Edu staining and colony formation assays. By performing loss-of-function and rescue experiments, we explored the role of HSRL and Sorting nexin 9 (SNX9) in TGF-β1-stimulated fibroblast. Employing RNA-protein pull-down assay and Co-immunoprecipitation, we further investigated the mechanisms through which TSIIA attenuated TGF-β1-stimulated fibroblast.

Results: Our data demonstrated that TSIIA could effectively attenuate TGF-β1-mediated fibroblast proliferation in a dose-dependent manner. Meanwhile, TSIIA could down-regulate the expression of α-SMA, VEGFA, Collagen 1, HSRL, SNX9 and p-Smad2/3 in TGF-β1-stimulated HSF. In addition, we found that SNX9 overexpression reversed the effects of HSRL knockdown on TGF-β1-stimulated HSF. Furthermore, we confirmed that TSIIA treatment weakens the interaction between p-Smad3 and SNX9 in HS models.

Conclusions: Tanshinone IIA down-regulated p-Smad3 signaling to inhibit TGF-β1-mediated fibroblast proliferation via lncRNA-HSRL/SNX9.

Keywords: Fibroblast; HSRL; Proliferation; SNX9; Tanshinone IIA; p-Smad3.