米诺环素通过Wnt/β-Catenin通路对大鼠牙髓干细胞成骨分化的影响

摘要/Abstract

摘要： 目的探讨米诺环素对大鼠牙髓干细胞（RDPSCs）成骨分化的影响，以及对Wnt/β-连环蛋白（Wnt/β-Catenin）通路的作用。方法将RDPSCs随机分为对照组和不同浓度（0.1、1.0、10.0、100.0 mg/L）米诺环素组，对细胞进行成骨诱导。CCK-8法检测细胞增殖率，试剂盒检测骨形成指标碱性磷酸酶（ALP）和骨钙蛋白（OCN）含量；茜素红S染色检测细胞钙盐沉积量；qRT-PCR检测ALP和OCN mRNA表达情况；蛋白质免疫印迹技术（WB）检测β-Catenin、糖原合成酶激酶-3β（GSK-3β）和Runt相关转录因子2（Runx2）蛋白表达水平。结果与对照组相比，0.1、1.0 mg/L米诺环素组RDPSCs细胞增殖率、ALP活性、OCN含量、钙盐沉积量及β-Catenin、Runx2蛋白水平均显著升高，GSK-3β蛋白水平显著降低（P＜0.05）；10.0、100.0 mg/L米诺环素组RDPSCs细胞增殖率显著升高（P>0.05），ALP活性、OCN含量及表达水平、钙盐沉积量及β-Catenin、Runx2蛋白水平均显著降低，GSK-3β蛋白水平显著升高（P＜0.05）。结论低浓度米诺环素能够诱导大鼠牙髓干细胞成骨分化，而高浓度米诺环素抑制细胞成骨分化，该过程可能通过调控Wnt/β-Catenin信号通路实现。

关键词: 米诺环素, Wnt/β-Catenin通路, 牙髓干细胞, 成骨分化, 碱性磷酸酶

Abstract: Objective To investigate the influence of minocycline on osteogenic differentiation of rat dental pulp stem cells (RDPSCs) and the effect on Wnt/β-Catenin pathway. Methods RDPSCs were randomly divided into control group and minocycline groups with different concentrations (0.1, 1.0, 10.0, 100.0 mg/L), and osteogenic differentiation was induced. CCK-8 method was used to detect the cell proliferation rate; the kit was used to detect alkaline phosphatase (ALP) and osteocalcin (OCN); alizarin red S staining was used to detect calcium deposition; qRT-PCR was used to detect ALP and OCN mRNA expression; Western blot (WB) was used to detect protein expression levels of β-Catenin, glycogen synthase kinase-3β (GSK-3β) and Runt related transcription factor 2 (Runx2). Results Compared with those in the control group, the proliferation rate, activity of ALP, content of OCN, calcium deposition, protein levels of β-Catenin and Runx2 were significantly increased in 0.1 and 1.0 mg/L minocycline groups; the protein level of GSK-3β was significantly decreased (all P＜0.05). The proliferation rate of RDPSCs in 10.0, 100.0 mg/L minocycline groups was significantly increased (P>0.05); activity of ALP, content of OCN and their expression levels, calcium deposition, protein levels of β-Catenin and Runx2 were significantly decreased; the protein level of GSK-3β was significantly increased (all P＜0.05). Conclusion Low-concentration minocycline can induce osteogenic differentiation of rat dental pulp stem cells, while high-concentration minocycline can inhibit osteogenic differentiation of rat dental pulp stem cells, which may be achieved by regulating Wnt/β-Catenin signaling pathway.

Key words: minocycline, Wnt/β-Catenin pathway, dental pulp stem cells, osteogenic differentiation, alkaline phosphatase

中图分类号:

R349.5

杜帅侠姚秀翠段珂王静. 米诺环素通过Wnt/β-Catenin通路对大鼠牙髓干细胞成骨分化的影响[J]. 口腔医学, 2021, 41(9): 776-781.

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