两种Wnt 通路抑制剂对牙周膜干细胞作用的比较研究

目的:本实验主要探讨Wnt通路抑制剂XAV-939相比DKK1在牙周膜干细胞增殖及矿化中的作用差异。方法:酶消化法培养牙周膜干细胞,鉴定后,用CCK8试剂盒检测XAV-939和DKK1对牙周膜干细胞增殖能力的影响,茜素红染色及定量检测XAV-939和DKK1对牙周膜干细胞成骨分化能力的影响,q RT-PCR检测DKK1和XAV-939对牙周膜干细胞Wnt通路相关基因GSK-3β和β-catenin及成骨分化相关基因ALP,DSPP,BSP,OCN,RUNX-2的影响。结果:XAV-939和DKK1都可以通过抑制Wnt通路来抑制牙周膜干细胞的增殖及成骨分化。当没有外源性Wnt蛋白刺激时,XAV-939作为Wnt通路抑制剂的抑制作用要强于DKK1,而加入外源性Wnt蛋白后,XAV-939与DKK1的作用效果相当。结论:XAV-939对比DKK1,具有更为广泛而稳定的抑制效果。XAV-939可以作为高效的Wnt通路抑制剂应用于未来关于牙周膜干细胞和Wnt通路相关实验研究中。

Comparative Research on the Effect of two Wnt Signaling Inhibitor on the Proliferation and Osteogenic Differentiation of PDLSC

Objective:To evaluate the effect of Wnt inhibitor XAV-939 on the proliferation and osteogenic differentiation of PDLSC and DKK1.Methods:PDLSCs were isolated from healthy parodontium of premolar teeth of individuals undergoing tooth extraction for orthodontic treatment. After identification, we used cck8 kit to evaluate the effect of XAV-939 and DKK1 on the proliferation of PDLSC. Alizarin Red Staining and quantification was used to evaluate the effect of XAV-939 and DKK1 on the osteogenic differentiation of PDLSC. The effect of XAV-939 and DKK1 on the gene expression of PDLSC, including Wnt signaling marker genes GSK-3beta, beta-catenin and osteogenic differentiation marker genes ALP, DSPP, BSP, OCN, RUNX-2 was evaluated by qRT-PCR.Results:XAV-939 and DKK1 could inhibit the proliferation and osteogenic differentiation of PDLSC by inhibiting Wnt passway. The inhibiting effect of XAV-939 was stronger than DKK1 when there wasn''t exogenous Wnt protein. And there weren''t statistical differences between XAV-939 and DKK1 after the addition of exogenous Wnt protein.Conclusion:The effect of XAV-939 was more extensive and stable. XAV-939 can be used as a high-efficiency Wnt inhibitor in the future research on PDLSC and Wnt passway.