Yin Lifen, Wang Xiao
Department of Periodontal Mucosa, Changsha Stomatological Hospital, Changsha, Hunan, 410004, P.R. China.
Cell Biochem Biophys. 2025 Jun;83(2):2227-2236. doi: 10.1007/s12013-024-01633-x. Epub 2024 Dec 15.
Oral submucous fibrosis (OSF) is a precancerous lesion of the oral cavity. Areca nut consumption can cause OSF through sustained activation of buccal mucosal fibroblasts (BMFs). This study explored the effect of curcumin on arecoline-induced BMF activation and its mechanism of action. BMFs were isolated and identified by immunofluorescence detection of fibroblast surface markers vimentin and S100A4. After transfection with FOSL1- or MAPK8-related vectors, BMFs were activated by arecoline and treated with curcumin. Scratch and transwell assays were performed to detect cell migration. ChIP and luciferase reporter assays were conducted to detect the binding of FOSL1 to the MAPK8 promoter. RT-qPCR was used to detect FOSL1 and MAPK8 mRNA expression. Western blotting was used to detect FOSL1, MAPK8, COL1A1, α-SMA, Smad2, and p-Smad2 proteins. Curcumin treatment inhibited arecoline-induced fibroblast migration, reduced the expression of myofibroblast markers COL1A1, α-SMA, and p-Smad2, and downregulated the expression of FOSL1 and MAPK8. FOSL1 or MAPK8 overexpression enhanced migration and increased COL1A1, α-SMA, and p-Smad2 expression in curcumin-treated cells. FOSL1 bound to the MAPK8 promoter and promoted MAPK8 expression. Simultaneous FOSL1 overexpression and MAPK8 knockdown, compared to FOSL1 overexpression, reduced cell migration and inhibited COL1A1, α-SMA, and p-Smad2 expression. In conclusion, curcumin targets FOSL1 to reduce MAPK8 expression, thereby suppressing arecoline-induced fibroblast activation.
口腔黏膜下纤维化（OSF）是口腔的一种癌前病变。食用槟榔可通过持续激活颊黏膜成纤维细胞（BMF）导致OSF。本研究探讨了姜黄素对槟榔碱诱导的BMF激活的影响及其作用机制。通过免疫荧光检测成纤维细胞表面标志物波形蛋白和S100A4分离并鉴定BMF。用FOSL1或MAPK8相关载体转染后，BMF用槟榔碱激活并用姜黄素处理。进行划痕试验和Transwell试验以检测细胞迁移。进行染色质免疫沉淀（ChIP）和荧光素酶报告基因试验以检测FOSL1与MAPK8启动子的结合。逆转录定量聚合酶链反应（RT-qPCR）用于检测FOSL1和MAPK8 mRNA表达。蛋白质免疫印迹法用于检测FOSL1、MAPK8、I型胶原蛋白α1（COL1A1）、α-平滑肌肌动蛋白（α-SMA）、Smad2和磷酸化Smad2蛋白。姜黄素处理抑制了槟榔碱诱导的成纤维细胞迁移，降低了肌成纤维细胞标志物COL1A1、α-SMA和磷酸化Smad2的表达，并下调了FOSL1和MAPK8的表达。FOSL1或MAPK8过表达增强了姜黄素处理细胞的迁移能力，并增加了COL1A1、α-SMA和磷酸化Smad2的表达。FOSL1与MAPK8启动子结合并促进MAPK8表达。与FOSL1过表达相比，同时过表达FOSL1和敲低MAPK8可减少细胞迁移并抑制COL1A1、α-SMA和磷酸化Smad2的表达。总之，姜黄素以FOSL1为靶点降低MAPK8表达，从而抑制槟榔碱诱导的成纤维细胞激活。
