Arecoline Promotes Fbroblast Activation And M2-Macrophage Polarization

To determine the biological effects of arecoline on oral submucous fbrosis (OSF). The differential genes between OSF tissue and normal oral tissue were collected form GSE64216 dataset, analyzed by Gene Expression Omnibus database. Real-time PCR and immunohistochemistry were used to analyze the expression of IL-4 in oral tissue. Enzyme-linked immunosorbent assay was used to analyze the expression of exocrine IL-4 protein in human oral fbroblasts (HOF) pre-treated by arecoline. Cell Counting Kit-8 and transwell assays were used to analyze the proliferation and migration of HOF cells, respectively. After IL-4 knocked down, the proliferation and migration of HOF cells were detected. Flow cytometry was used to analyze the proportion of M2-macrophages. Real-time PCR and immunohistochemistry were used to verify the expression of biomarker proteins of macrophages in OSF tissues. The expression of IL-4 was up-regulated in OSF tissue .Arecoline could enhance the expression of IL-4 gene and protein in HOF cells, and promote the proliferation and migration of HOF cells. While knockdown of IL-4 could inhibit arecoline-induced proliferation and migration in HOF cells. The recombinant human IL-4 (rhIL-4) protein could increase the pro-portion of M2-macrophages. Similarly, the results of real-time PCR and immunohistochemistry showed the expression of ARG1 (Biomarker proteins of M2-macrophage) was up-regulated in OSF tissues. Arecoline promotes activation of fbroblasts and polarization of M2-macrophages by up-regulating the expression of IL-4.