Changsha Stomatological Hospital, Changsha 410000, China.
Xiangya Stomatological Hospital, Central South University, Changsha 410000, China.
Arch Oral Biol. 2024 Nov;167:106052. doi: 10.1016/j.archoralbio.2024.106052. Epub 2024 Jul 20.
To determine the biological effects of arecoline on oral submucous fibrosis (OSF).
The differential genes between OSF tissue and normal oral tissue were collected form GSE64216 dataset, analyzed by Gene Expression Omnibus (GEO) database. Real-time PCR and immunohistochemistry were used to analyze the expression of IL-4 gene and protein in oral tissue. Enzyme-linked immunosorbent assay (ELISA) was used to analyze the expression of exocrine IL-4 protein in human oral fibroblasts (HOF) pre-treated by arecoline. Cell Counting Kit-8 (CCK-8) and transwell assays were used to analyze the proliferation and migration of HOF cells, respectively. After IL-4 was knocked down by short hairpin (sh) plasmid, 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 gene and protein were both up-regulated in OSF tissue. Arecoline could enhance the expression of IL-4 gene and exocrine 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 results of flow cytometry showed that recombinant human IL-4 (rhIL-4) protein could increase the proportion 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 fibroblasts and polarization of M2-macrophages by up-regulating the expression of IL-4.
研究槟榔碱对口腔黏膜下纤维化（OSF）的生物学作用。
从 GSE64216 数据集收集 OSF 组织与正常口腔组织的差异基因，通过基因表达综合数据库（GEO）进行分析。采用实时 PCR 和免疫组织化学法分析口腔组织中白细胞介素 4（IL-4）基因和蛋白的表达。采用酶联免疫吸附试验（ELISA）分析槟榔碱预处理人口腔成纤维细胞（HOF）中外分泌型 IL-4 蛋白的表达。采用细胞计数试剂盒 8（CCK-8）和 Transwell 检测分别分析 HOF 细胞的增殖和迁移。用短发夹（sh）质粒敲低 IL-4 后，检测 HOF 细胞的增殖和迁移。采用流式细胞术分析 M2-巨噬细胞的比例。采用实时 PCR 和免疫组织化学法验证 OSF 组织中巨噬细胞生物标志物蛋白的表达。
OSF 组织中 IL-4 基因和蛋白表达均上调。槟榔碱可增强 HOF 细胞中 IL-4 基因和外分泌蛋白的表达，并促进 HOF 细胞的增殖和迁移。而敲低 IL-4 可抑制 HOF 细胞中槟榔碱诱导的增殖和迁移。流式细胞术结果表明，重组人白细胞介素 4（rhIL-4）蛋白可增加 M2-巨噬细胞的比例。同样，实时 PCR 和免疫组织化学法结果表明，OSF 组织中 ARG1（M2-巨噬细胞的生物标志物蛋白）表达上调。
槟榔碱通过上调 IL-4 的表达，促进成纤维细胞的激活和 M2-巨噬细胞的极化。
