力生长因子调控牙周膜干细胞Src-YAP/TAZ通路促牙周膜再生修复的分子机制研究

本课题组前期研究发现，力生长因子MGF是力学刺激促牙周膜干细胞向牙周膜成纤维细胞分化进而促牙周膜再生修复的重要信号分子；Src家族的Lyn、Fyn 及转录辅助因子YAP/TAZ参与该调控过程，但其具体信号转导通路尚有待进一步深入研究。本项目将综合利用RNAi、基因转染等方法检测MGF作用下整合素、FAK、G蛋白等Scr相关已知细胞膜力学感受蛋白的响应；结合计算机蛋白预测与GST-pull down技术对细胞膜中可能存在的MGF特异性受体蛋白进行研究，以阐明MGF信号由细胞膜转导至细胞浆的分子机制。同时采用慢病毒转染、Co-IP检测等方法揭示YAP/TAZ在MGF促牙周膜再生修复中所起的转录调控作用，以阐明MGF信号由细胞浆转导至细胞核的分子机制。本课题将从体内外两方面揭示力生长因子调控牙周膜干细胞分化、进而促牙周膜再生修复的分子机理，为撕脱再植牙获得理想的牙周膜性愈合提供新的策略和思路。

Our previous study found that the mechano- growth factor (MGF) is an important signaling molecule for mechanical stimulation to stimulate the differentiation of periodontal ligament stem cells (PDLSCs) into periodontal ligament fibroblasts, thereby promoting the repair and regeneration of periodontal ligament. We also found that Lyn, Fyn and transcription of Src family The cofactor YAP/TAZ are involved in this regulation process, but its specific signal transduction pathway remains to be further studied. This project will comprehensively utilize RNAi and gene transfection methods to detect the response of known mechano-sensor proteins on the cell membrane relating to Scr family, including integrin, FAK, G protein, etc under the treatment of MGF. Combined with computer protein prediction and GST-pull down technology, MGF-specific receptor proteins that may exist in cell membrane of PDLSCs will be studied to elucidate the molecular mechanism by which MGF signals are transduced from the cell membrane to the cytosol. At the same time, lentiviral transfection and Co-IP detection will be used to reveal the transcriptional regulation of YAP/TAZ in MGF-promoting periodontal ligament regeneration, in order to elucidate the molecular mechanism of MGF signaling from cytoplasm to nucleus. This subject will reveal the molecular mechanism of MGF regulating the differentiation of PDLSCs and further promoting the regeneration of periodontal ligament from both in vitro and in vivo, and provide new strategies and ideas for obtaining a satisfactory periodontal healing for avulsed teeth after replantation.