Huaier cream attenuates TGF-β-induced keloid Fibroblast proliferation and migration by inhibiting the TGF-β/Smad signaling pathway

OBJECTIVE: To investigate whether Huaier cream (HC) regulates the Transforming growth factor-β/Smad signaling pathway (TGF-β/SSP) and influences the proliferation and migration (P&M), apoptosis, and the synthesis of extracellular matrix (ECM) in keloid fibroblasts (KFs). METHODS: The KFs model was established in vitro. Live/dead staining (L/D-S) and CCK-8 assays were performed to assess cell proliferation, while the wound healing assay (WHA) was used to evaluate cell migration. Flow cytometry (FC) was taken to assess cell apoptosis. Western blotting (WB) and immunofluorescence (IF) were conducted to determine the changes in TGF-β/SSP and analyze the expression of ECM-related Protein (ECM-RP). RESULTS: HC significantly inhibited the P&M of TGF-β-induced KFs (TGF-β-IKFs), restored their apoptotic capacity, and decreased the expression of ECM-RPs in a concentration-dependent manner. Additionally, HC treatment markedly lowered the Smad2/3's phosphorylation levels, indicating inhibition of the TGF-β/SSP. By suppressing the TGF-β/SSP, HC can inhibit the phosphorylation of TGF-β/SSP and regulate the P&M, apoptosis, and ECM synthesis of TGF-β-IKFs. CONCLUSIONS: HC suppresses the TGF-β/SSP, thereby inhibiting the P&M, and ECM synthesis of KFs, while inducing fibroblast apoptosis. These conclusions demonstrate that HC is a potential therapeutic agent for treating keloids.