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目的探讨氧联乙酰葡萄糖胺(O-linked N-acetylglucosamine,O-Glc NAc)影响上唇发育时上皮层的消失过程中,Wnt信号通路在其中发挥的作用。方法通过在鸡胚唇部植入浸有O-GlcNAc促进剂氧联乙酰葡萄糖胺转移酶(O-GlcNAc transferase,OGT)、O-GlcNAc的抑制酶氧联乙酰葡萄糖胺水解酶(O-Glc NAcase,OGA)以及OGA的抑制剂链脲佐菌素(streptozoticin,SZT)的琼脂糖微球,构建O-Glc NAc高表达与低表达以及反向高表达实验组,并继续孵育24 h,茜素红—阿利新蓝骨组织染色观察胚胎骨、软骨发生的变化,TUNEL法检测细胞凋亡情况,免疫组化染色法检测Wnt信号系包括GSK-3β、β-catenin在内的一系列信号因子。结果 O-GlcNAc过表达组OGT和STZ中,上皮层的消失出现延迟,骨染色发现胚胎唇部出现骨缺损,TUNEL检测与正常组相比未发现明显细胞凋亡,但GSK-3β在细胞质内表达增加,β-catenin细胞核内表达减少;正常侧及O-GlcNAc低表达组OGA中上唇发育未受影响。结论 O-GlcNAc过表达时引起上皮层的消失发生延迟,这并非由细胞凋亡减少引起,而是可能通过抑制Wnt信号通路,导致GSK-3β、β-catenin在内的相关信号因子变化,这些变化将影响上唇发育并可能导致唇腭裂的发生。
OBJECTIVE: To investigate the role of O-linked nicotinamide (O-GlcNAc) in the loss of epithelium during upper lip development. Methods O-GlcNAc transfected O-GlcNAc transferase (OGT) and O-GlcNAc-O-GlcNAcase , OGA), and OGA inhibitor streptozotocin (SZT). The O-GlcNAc high-expression and low-expression and reverse high expression group were constructed. The changes of embryonic bony and cartilage were observed by staining with red-Allee blue bone. The apoptosis was detected by TUNEL method. A series of signal factors including GSK-3β and β-catenin were detected by immunohistochemical staining. Results OGT and STZ in O-GlcNAc overexpression group showed delayed disappearance of epithelial layer. Bone defects were found in the lip of embryonic bone in O-GlcNAc overexpression group. TUNEL assay showed no significant apoptosis compared with normal group. However, GSK-3β was found in cytoplasm The expression of β-catenin was decreased in the nucleus and the upper lip of OGA was not affected in normal and O-GlcNAc low expression groups. Conclusions The overexpression of O-GlcNAc leads to the delay of the disappearance of epithelial layer, which is not caused by the decrease of apoptosis, but may be caused by the inhibition of Wnt signaling pathway and the related signal factors leading to GSK-3β and β-catenin Changes will affect the upper lip development and may lead to cleft lip and palate.