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

发布时间：2017-12-28 04:19

本文关键词：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　出处：《华西口腔医学杂志》2015年06期 　论文类型：期刊论文

【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(h PDLSCs)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h PDLSCs,实验分两组:正常h PDLSCs组(N-h PDLSCs组)和AGEs刺激的正常h PDLSCs组(A-h PDLSCs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-h PDLSCs组ALP染色明显浅于N-h PDLSCs组;茜素红染色A-h PDLSCs组钙化结节少于N-h PDLSCs组;real time PCR与Western blot的结果显示A-h PDLSCs组BSP、ALP、Runx-2的表达都较低,差异有统计学意义(P0.05)。Wnt经典信号通路中相关因子:A-h PDLSCs组β-catenin表达高于N-h PDLSCs组,A-h PDLSCs组DKK-1表达明显低于N-h PDLSCs组,并且Weston blot显示A-h PDLSCs组核蛋白β-catenin的表达高于N-h PDLSCs组。结论 AGEs可以使h PDLSCs骨向分化过程中Wnt经典通路的抑制因子DKK-1下调,细胞核中的β-catenin表达升高,激活了Wnt经典通路,并且抑制了骨分化。
[Abstract]:Objective to investigate the changes of Wnt classical pathway related factors Dickkopf protein 1 (DKK-1) and beta chain protein (beta -catenin) in the process of bone formation of human periodontal ligament stem cells (H PDLSCs) mediated by advanced glycation end products (AGEs). Methods h PDLSCs was cloned and cultured in vitro by tissue block method and limited dilution method. The experiment was divided into two groups: normal h PDLSCs group (N-h PDLSCs group) and AGEs stimulation normal h PDLSCs group (A-h PDLSCs group). 2 groups of cells of bone mineralization induced by alkaline phosphatase (ALP) and alizarin red staining; real-time polymerase chain reaction (real time PCR) detection of osteogenic gene, and related factors Wnt classic pathway of DKK-1, -catenin Western blot beta; detection of osteogenic protein and nuclear protein beta -catenin. Results after osteogenic induction, A-h ALP staining in PDLSCs group is obviously lighter than the N-h PDLSCs group A-h PDLSCs group; alizarin red staining of calcified nodules less than N-h group PDLSCs real time PCR Western blot; and the results showed that the expression of A-h PDLSCs BSP, ALP, Runx-2 group were lower, the difference was statistically significant (P0.05). The related factors in Wnt classic signal pathway: A-h PDLSCs group -catenin expression is higher than that in N-h PDLSCs group, DKK-1 expression in A-h PDLSCs group is significantly lower than that in N-h PDLSCs group, and the expression of nuclear protein in beta group is higher than that in N-h group. Conclusion AGEs can downregulate the inhibitory factor DKK-1 of Wnt classical pathway in the process of H PDLSCs bone marrow differentiation, and increase the expression of beta -catenin in the nucleus, activate Wnt classical pathway, and inhibit bone differentiation.
【作者单位】：遵义医学院附属口腔医院;贵阳市口腔医院牙周科;第四军医大学组织工程中心;
【基金】：国家自然科学基金资助项目(81360168)
【分类号】：R587.2;R781.4
【正文快照】： 糖基化终末产物(advanced glycation end pro-ducts,AGEs)与糖尿病多种并发症的发生密切相关[1]。近年来,有研究[2]表明糖尿病患者晚期AGEs的大量积聚可促进糖尿病的发展,并且可诱导牙周病的发生和发展。人牙周膜干细胞(human periodon-tal ligament stem cells,h PDLSCs)具有

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