O-GlcNAc对上唇发育过程中上皮间叶转化影响的研究

目的: 研究氧联乙酰葡萄糖胺(O-linked N-acetylglucosamine,O-GlcNAc)在上唇发育过程中对面突间上皮间叶转化的影响。方法: 通过在鸡胚上唇植入浸有O-GlcNAc促进剂 OGT(O-GlcNAc transferase)、O-GlcNAc 的抑制酶OGA(O-GlcNAcase)、以及OGA的抑制剂STZ(streptozoticin)的琼脂糖微球,构建O-GlcNAc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,免疫组化染色法检测具有间叶组织表达特异性的fibronectin和上皮表达特异性的p63的表达状态。结果: 高表达O-GlcNAc实验组OGT和STZ中,发现胚胎上唇出现了骨缺损,且P63、fibronectin持续高表达,上皮、间叶组织保持连续性,凋亡出现延迟,上皮间叶转化出现障碍。正常对照测与低表达OGA组在上皮融合过程中P63表达减弱,fibronectin在上皮中出现表达,上皮层出现断裂,表明上皮间叶转化的正常进行。 结论: O-GlcNAc过表达对于上唇形成过程中上皮间叶转化具有负面作用,将影响上唇的正常发育并可能导致唇裂的发生。

Influence of O-GlcNAc on the Epithelial Mesenchymal Transitios of Facial Process in the Development of Upper Lip.

Objective: To explore the influence of O-linked N-acetylglucosamine on the epithelial mesenchymal transition of facial process in the development of upper lip. Methods: By transplanting beads into the upper lip of chicken embryo which contained O-GlcNAc promoter OGT (O-GlcNAc transferase), O-GlcNAc inhibitor OGA (O-GlcNAcase) or OGA inhibitor STZ (streptozoticin), the over/lower-expression of O-GlcNAc chicken were constructed. After 24h prototypical analysis of skeletal and morphological changes was performed using Alizarin red/Alcian blue method. Fibronectin and P63 were detected by immunohistochemistry method. Results: Overexpression of O-GlcNAc led to bone defect in the area of upper lip, and P63 and fibronectin were detected frequently constitutively activated during the development of upper lip, which led to delay and obstacle of epithelial-mesenchymal transition. While there were no serious abnormalities noted on the normal group and the lower-expression group. Conclusion: Overexpression of O-GlcNAc may lead to delay and obstacle of epithelial-mesenchymal transition, which may influence the development of upper lip and may lead to cleft lip.