糖基化终末产物对牙周膜干细胞增殖及其Wnt经典信号通路相关基因表达的影响

目的:观察糖基化终末产物(AGEs)对人牙周膜干细胞(HPDLSC)增殖及其Wnt经典信号通路相关基因DKK-1、β-catenin表达的影响。方法:体外组织块法和有限稀释法克隆化培养HPDLSC,并与100μg/mL AGEs共同培养,通过细胞克隆形成率、细胞生长曲线观察AGEs对HPDLSC增殖能力的影响;RT-PCR和Western Blot检测实验组和对照组DKK-1、β-catenin mRNA以及核蛋白β-catenin的表达量。结果:经100μg/mL AGEs刺激的HPDLSC,其克隆形成率、增殖速度以及β-catenin mRNA和核蛋白β-catenin的表达量均明显低于对照组(P<0.05);而DKK-1 mRNA则明显高于对照组(P<0.05)。结论:AGEs能抑制HPDLSC的增殖及其Wnt经典信号通路。

Effects of advanced glycation end products on proliferation of human periodontal ligament stem cells and Wnt signaling

AIM: To investigate the effects of advanced glycation end products(AGEs) on proliferation of human periodontal ligament stem cells(HPDLSCs) and expressions of DKK-1 and β-catenin,genes of Wnt signaling pathway.METHODS: PDLSCs were isolated and cultured by limited dilution method to single clones.Flow cytometry was employed to study the surface markers of the cloned cells.Single clones of PDLSCs were stimulated with 100 g/mL of AGEs.Proliferation of PDLSCs was measured by MTT assay.After 7 days stimulation,mRNA and nuclear proteins were extracted to determined the gene and protein expression of beta catenin and DKK-1 by real-time quantitative polymerase chain reaction(real time PCR) and Westen blot.RESULTS:Clone formation rates of unstimulated and stimulated PDLSCs were 1.6% and 17.2%,respectively(P<0.05).The proliferation rate of stimulated PDLSCs was significantly lower than that of the mstimulatad group by MTT assay.The mRNA and protein expression of beta catenin was significantly higher in unstimulated PDLSCs,whereas DKK-1 expression was significantly lower in unstimulated PDLSCs as compared with AGEs stimulated cells(P﹤0.05).CONCLUSION: AGEs may inhibit the proliferation of human periodontal ligament stem cells by suppressing the gene expression of Wnt signaling pathways.