EOFAZ inhibits endothelial-to-mesenchymal transition through downregulation of KLF4

Abstract

Essential oil from Alpinia zerumbet rhizome (EOFAZ), which is termed Yan shanjiang in China, is extensively used as an herbal medicine in the Guizhou area and has been shown to protect against the damaging effects of cardiovascular injury in vitro and in vivo. In the present study, it was hypothesized that the protective effects of EOFAZ on transforming growth factor (TGF)-β1-induced endothelial-to-mesenchymal transition (EndMT) in human umbilical vein endothelial cells (HUVECs) were mediated by inhibition of Kruppel-like factor 4 (KLF4). Cell motility was assessed using wound healing and Transwell assays. The expression of endothelial markers and mesenchymal markers were determined by reverse transcription-quantitative PCR, immunofluorescence staining and western blotting, and additionally, phosphorylated NF-κB p65 expression was determined by western blotting. Furthermore, the involvement of KLF4 in EndMT was determined using RNA interference to knockdown the expression of KLF4. TGF-β1 treatment significantly promoted EndMT, as evidenced by downregulation of vascular endothelial-cadherin and upregulation of α-smooth muscle actin in HUVECs, and by enhancing cell migration. Small interfering RNA-mediated knockdown of KLF4 reversed TGF-β1-induced EndMT. Additionally, treatment with EOFAZ inhibited TGF-β1-induced EndMT in a dose-dependent manner. These results suggest that TGF-β1 may induce EndMT through upregulation of KLF4, and this may be reversed by EOFAZ. Therefore, EOFAZ was shown to inhibit TGF-β1-induced EndMT through regulation of KLF4.