褪黑素经Wnt/β-catenin信号通路调控hDPSCs的成骨分化
Melatonin regulates osteogenic differentiation of hDPSCs through Wnt/β-catenin signaling pathway
李婉昕 1李思美 1潘宏伟 1王佳 1殷丽华1
作者信息
- 1. 730000,兰州大学口腔医学院(口腔医院)口腔种植科
- 折叠
摘要
目的:探究褪黑素对人牙髓干细胞(hDPSCs)增殖及成骨分化的调控作用及相关机制.方法:体外分离培养hDPSCs,空白对照组细胞常规培养,实验组细胞分别使用成骨诱导培养基(OIM)和不同浓度(5、10、25、50、100、250、500和1 000 µmol/L)褪黑素培养.CCK-8实验检测褪黑素对hDPSCs增殖能力的影响,检测各组细胞碱性磷酸酶(ALP)活性,RT-qPCR、Western blot分析成骨分化相关mRNA及蛋白表达水平以及Wnt通路相关蛋白表达水平.结果:褪黑素浓度为100 μmol/L和250 µmol/L时可促进hDPSCs增殖;其中,100 µmol/L浓度条件下hDPSCs各成骨相关mRNA和蛋白表达水平均明显上升,并上调β-catenin、下调GSK-3β表达水平.结论:特定浓度范围内褪黑素可促进hDPSCs增殖,且具备调节Wnt/β-catenin信号通路以促进hDPSCs成骨分化的潜能.
Abstract
Objective:To explore the regulatory effects and mechanism of melatonin on the proliferation and osteogenic differentiation of human dental pulp stem cells(hDPSCs).Methods:hDPSCs were cultured in vitro.The cells in the blank control groups were rou-tinely cultured,while the cells in the experimental groups were cultured with osteogenic induction medium(OIM)and OIM with me-latonin at 5,1,10,25,50,100,500 and 1 000 µmol/L respectively.The effect of melatonin on the proliferation of hDPSCs was ob-served by CCK-8 assay.Alkaline phosphatase(ALP)test,RT-qPCR and Western blot were used to analyze the expression of osteo-genic related genes and proteins.The expression level of the proteins related to classical Wnt pathway was detected.Results:Melato-nin at 100 and 250 µmol/L promoted the proliferation of hDPSCs,at 100 μmol/L increased the expression levels of osteogenesis-related mRNA and proteins in hDPSCs,up-regulated β-catenin and down-regulated GSK-3β expression.Conclusion:Melatonin at specific concentrations can promote the proliferation of hDPSCs and has the potential to regulate Wnt/β-catenin signal pathway to pro-mote osteogenic differentiation of hDPSCs.
关键词
褪黑素/牙髓干细胞/增殖/成骨分化Key words
Melatonin/hDPSCs/Proliferation/Osteogenic differentiation引用本文复制引用
基金项目
甘肃省省级引导科技创新发展竞争性项目(2018ZX-10)
甘肃省科技计划国际合作专项(18YF1WA116)
兰州市人才创新创业项目(2017-RC-31)
口腔医学科研扶持基金团队项目(lzukqky-2019-t11)
出版年
2024
NETL
NSTL
万方数据