中文摘要：

目的：研究阻断经典Wnt通路对棕色脂肪干细胞向起搏样细胞分化的影响.方法：在诱导棕色脂肪干细胞向起搏特性诱导的过程中,培养基中加入经典Wnt通路阻断剂Dkk-1培养10 d.相差显微镜观察诱导组细胞的形态学变化、应用real-time PCR检测其中起搏细胞发育相关结构基因的表达情况,免疫荧光显色普通光镜和激光共聚焦显微镜观察起搏特征分子的蛋白表达.结果：分化后的棕色脂肪干细胞具有起搏细胞特性,经Dkk-1诱导后,棕色脂肪干细胞向起搏样细胞的转化率明显提高,起搏细胞发育相关基因和蛋白表达明显上调.结论：阻断经典Wnt通路可促进棕色脂肪干细胞向起搏样细胞的分化.

英文摘要：

Objective： To study the role of inhibition of canonical Wnt pathway in differentiation of brown adipose-derived stem cells （BADSCs） into pacemaker-like cells. Methods： Dkk-1, a canonical Wnt antagonist, was added into the medium for 10 days when the BADSCs were induced into pacemaker-like cells. The morphological variations after induction were observed by phase contrast microscope. The expression of genes related to the development of the sinoatrial node were measured by real- time PCR, while the expression of pacing-related proteins were stained by immunofluorescence and detected by regular and confocal laser microscope. Results： Differentiated BADSCs presented properties of pacemaker-like cells. In the presence of Dkk-1, BADSCs could differentiate into more spontaneously beating cells, and expressed more pacing-related genes and proteins. Conclusion ： Inhibition of canonical Wnt signal pathway could promote the differentiation of BADSCs into pacemaker- like cells.