Tideglusib对LPS刺激的人根尖牙乳头干细胞牙/骨向分化的影响

doi:10.13591/j.cnki.kqyx.2022.07.004

摘要/Abstract

摘要： 目的探讨Tideglusib对LPS刺激的人根尖牙乳头干细胞(stem cells from the apical papilla, SCAPs)的牙/骨向分化的影响。方法分离培养SCAPs,流式细胞术对SCAPs进行表面分子鉴定,检测Tideglusib对SCAPs细胞增殖是否有影响。通过碱性磷酸酶(alkaline phosphatase, ALP)活性和ALP染色筛选Tideglusib促进SCAPs表达ALP活性的最佳浓度。用大肠杆菌脂多糖(lipopolysaccharide, LPS)模拟炎性微环境刺激SCAPs。采用Western blot及实时定量聚合酶链反应(quantitative real-time polymerase chain reaction, RT-qPCR)等方法检测牙/骨向分化的相关蛋白(OSX、OCN、COL-Ⅰ、DSP、RUNX2)和基因(OSX、OCN、COL-Ⅰ、DSPP、RUNX2)表达变化。结果 CCK-8实验显示:Tideglusib浓度低于50 nmol/L时对细胞增殖无抑制作用;1 nmol/L Tideglusib处理LPS刺激的SCAPs,其ALP活性增加最明显;Western blot及qRT-PCR实验显示:1 nmol/L Tideglusib处理后的LPS刺激的SCAPs其成骨/成牙相关蛋白(OSX、OCN、COL-Ⅰ、DSP、RUNX2)和基因的表达(OSX、OCN、COL-Ⅰ、DSPP、RUNX2)均明显上调(P<0.05)。结论 1 nmol/L Tideglusib可以促进LPS刺激的SCAPs的牙/骨向分化能力。

关键词: Tideglusib, 脂多糖, 根尖牙乳头干细胞, 分化

Abstract: Objective To investigate effects of Tideglusib on the osteo/odontogenic differentiation of stem cells from the apical papilla stimulated by lipopolysaccharide(LPS). Methods Human stem cells from the apical papilla (SCAPs) were isolated and cultured from human dental papilla, and the surface molecules were identified by flow cytometry. CCK-8 kit was used to detect whether Tideglusib affected cell proliferation of SCAPs. The optimal concentration of Tideglusib promoting ALP activity of SCAPs was screened by alkaline phosphatase (ALP) activity and ALP staining. LPS of Escherichia coli were used to create an inflammatory microenvironment. Western blot and RT-qPCR were applied to investigate changes of proteins (OSX, OCN, COL-Ⅰ, DSP, RUNX2) and genes (OSX, OCN, COL-Ⅰ, DSPP, RUNX2) associated with osteo/odontogenic differentiation. Results CCK-8 results showed that Tideglusib with the concentration below 50 nmol/L had no cytotoxin (P>0.05); ALP results showed that the ALP activity of SCAPs stimulated by LPS treated with 1 nmol/L Tideglusib increased the most significantly (P<0.05); Western blot and RT-qPCR showed that the expression of osteo/odontogenic related proteins (OSX、OCN、COL-Ⅰ、DSP、RUNX2) and genes (OSX, OCN, COL-Ⅰ, DSPP, RUNX2) of SCAPs stimulated by LPS treated with 1 nmol/L Tideglusib markedly increased (P<0.05). Conclusion Tideglusib (1 nmol/L) can promote the osteo/odontogenic differentiation ability of SCAPs stimulated by LPS.

Key words: Tideglusib, LPS, stem cells from the apical papilla, differentiation

中图分类号:

R780.2

蒋玫, 张悦蓉, 沈天晖, 张光东. Tideglusib对LPS刺激的人根尖牙乳头干细胞牙/骨向分化的影响[J]. 口腔医学, 2022, 42(7): 593-599.

JIANG Mei, ZHANG Yuerong, SHEN Tianhui, ZHANG Guangdong. Effects of Tideglusib onosteo/odontogenic differentiation of human stem cells from the apical papilla stimulated by LPS[J]. Stomatology, 2022, 42(7): 593-599.

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