The correlation between TGF-b3 and HGF/SF in the process of fusion of secondary palatal shelves

本研究在体外和体内研究了调节胚胎期细胞生长的TGF-β与器官培养中二级腭架融合过程中上皮-间质转化相关的HGF/SF的相关机制。胚胎第12 ~ 15天采用H.E.染色法检测腭发育，第12 ~ 15天采用全载原位杂交法检测HGF/SF和TGF-β3 Mrna的表达。此外，我们还研究了HGF/SF或TGF-β3在器官培养中的作用。TGF-β3 Mrna从胚胎第12天开始在上皮和间充质层中表达，在胚胎第12.5天显著表达，在胚胎第13.5天后消失；HGF/SF Mrna从胚胎第12天开始在间充质层中表达，在胚胎第13.5天后消失。HGF/SF反义寡核苷酸处理可抑制小鼠二级腭架的融合，但不抑制其在器官培养中的发育。c-Met中和抗体处理可抑制小鼠器官培养中二级腭架的发育。TGF-β3反义寡核苷酸处理可抑制器官培养小鼠二级腭架的发育和融合。上述结果提示，HGF/SF和TGF-β3可能在同一胚胎阶段表达，HGF/SF可能在小鼠二级腭架由垂直方向向水平方向变化时诱导腭尖细胞凋亡，TGF-β3可能与小鼠二级腭架向水平方向生长有关。

In this study, we investigated the mechanism of the corrrelation between TGF-β which regulates the cell growth in the embryonic stage anf HGF/SF which is related to epithelial-mesenchymal transformation in the process of the fusion of secondary palatal shelves in organ culture in vitro and vivo.Palate development was examined by H.E. staining on embryonic day 12〜15, and expression of HGF/SF and TGF-β3, Mrna was investigated by whole mount in situ hybridisation on embryonic day 12〜15. In addition, we conducted to examine whether treatment of HGF/SF or TGF-β3 in organ culture.Expression of TGF-β3 Mrna was detected in epithelial and mesenchymal layer from embryonic day 12, and they were observed markedly on embryonic day 12.5, and disppeared after embryonic day 13.5, Expression of HGF/SF Mrna was also detected in mesenchymal layer from embryonic day 12, and disppeared after embryonic day 13.5. Treatment of HGF/SF antisense oligonucleotide prove inhibited the fusion of mouse secondary palatal shelves,but did not inhibit the development of them in organ culture. Treatment of c-Met neutralizing antibody inhibited the development of mouse secondary palatal shelves in organ culture. Treatment of TGF-β3 antisense oligonucleotide prove inhibited the development and fusion of mouse secondary palatal shelves in organ culture.These findings suggested that HGF/SF and TGF-β3 may express at the same embryonic stage, and HGF/SF may induce apoptosis in the tip of the palate at the time which mouse secondary palatal shelves make a change of direction from vertical position to horizontal position, and TGF-β3 may be related with the growth of mouse secondary palatal shelves to the horizontal direction.