糖基化终末产物对人牙周膜干细胞骨向分化过程中的Wnt经典信号通路研究
Canonical Wnt signaling pathway of the osteogenic differentiation of human periodontal ligament stem cells induced by advanced glycation end products
作者：伍燕,邓超,杨琨,崔晓霞,刘琪,金岩
Author：Wu Yan, Deng Chao, Yang Kun, Cui Xiaoxia, Liu Qi, Jin Yan
收稿日期：2015-05-11 年卷（期）页码：2015,33(6):627-627-632
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：糖基化终末产物,人牙周膜干细胞,骨向分化,Wnt经典通路,
Key words：advanced glycation end products,human periodontal ligament stem cells,osteogenic differentiation,classical Wnt pathway,
基金项目：国家自然科学基金资助项目（81360168）
中文摘要
目的 探讨Wnt经典通路相关因子Dickkopf蛋白1（DKK-1）、β-链蛋白（β-catenin）在糖基化终末产物（AGEs）介导的人牙周膜干细胞（hPDLSCs）骨分化过程中的变化。方法 通过体外组织块法和有限稀释法克隆化培养获取hPDLSCs，实验分两组：正常hPDLSCs组（N-hPDLSCs组）和AGEs刺激的正常hPDLSCs组（A-hPDLSCs组）。对2组细胞成骨矿化诱导后行碱性磷酸酶（ALP）和茜素红染色；实时聚合酶链反应（real time PCR）检测成骨基因，以及Wnt经典通路中相关因子DKK-1、β-catenin；Western blot检测相关成骨蛋白、细胞核蛋白β-catenin。结果 成骨诱导后，A-hPDLSCs组ALP染色明显浅于N-hPDLSCs组；茜素红染色A-hPDLSCs组钙化结节少于N-hPDLSCs组；real time PCR与Western blot的结果显示A-hPDLSCs组BSP、ALP、Runx-2的表达都较低，差异有统计学意义（P
英文摘要
Objective The effect of advanced glycation end products (AGEs) on the osteogenic differentiation of human periodontal ligament stem cells(hPDLSCs) was discussed. Changes in the Wnt signaling pathway during glycation were also determined. MethodsIn vitrotissue explanting method was primarily applied. Limiting diluted clone was cultured to obtain hPDLSCsin vitro. The subjects were divided into two groups: the healthy group (N-hPDLSCs) and the AGEs-stimulating group (A-hPDLSCs). Osteoblast mineralization was induced in the experimental groups. The following processes were performed: alizarin red staining; alkaline phosphatase (ALP) staining; real time polymerase chain reaction (real time PCR) for detecting osteogenic genes and Wnt classical pathway-related factors, DKK-1 and β-catenin; Western blot analysis. Bone protein and β-catenin were correlated in the nuclear expression. Results The cells were osteogenically induced. ALP staining showed that the N-hPDLSCs displayed the deepest color. Alizarin red staining indicated that the A-hPDLSCs group had less calcified nodules than the N-hPDLSCs group. The real time PCR results suggested that the expression of relative osteogenic genes in A-hPDLSCs was quite low. Statistically significant differences in differentiation were found between groups (P
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