维生素K2促进BM-MSCs成骨分化及其机制研究
Vitamin K2 Promotes Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells via Runx2 Signaling Pathway
  
DOI:10.3969/j.issn.1006-7108.2020.06.007
中文关键词:  维生素K2  BM-MSCs  成骨分化  BMP-2
英文关键词:vitamin K2  BM-MSCs  osteogenic differentiation  BMP-2
基金项目:福建省卫生健康委员会青年课题(2012-2-5)
作者单位
许志贤 何武兵 柯铁 张永发 傅超锋 蔡加琴 张桂枫* 福建医科大学省立临床医学院/福建省立医院,福建 福州 350001 
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中文摘要:
      目的 探讨维生素K2对骨髓来源间充质干细胞(BM-MSCs)成骨分化能力的影响和机制。方法 通过CCK-8 活细胞计数法检测维生素K2不同浓度(1 nmol/L ,10 nmol/L, 100 nmol/L, 1 μmol/L,10 μmol/L ) 在不同时间( 24、48、72 h) 对BM-MSCs生长增殖的影响,运用Q-PCR检测不同浓度维生素K2对BM-MSCs成骨分化相关基因BMP-2表达水平的影响,进一步探讨维生素K2是否通过成骨信号通路Smad1/5/8、Runx2及Osterix调控BM-MSCs成骨分化。结果 与对照组比较,低浓度(1 nmol/L)的维生素K2不影响BM-MSCs的生长活性,中浓度(10 nmol/L, 100 nmol/L, 1 μmol/L)明显促进BM-MSCs的生长,高浓度(10 μmol/L)明显抑制细胞的生长(P<0.05)。中浓度范围的维生素K2呈剂量依赖性地促进BM-MSCs成骨过程中BMP-2 mRNA表达及钙结节的形成,且1 μmol/L的维生素K2明显促进BM-MSCs的Smad1/5/8、Runx2 及 Osterix 蛋白水平。结论 维生素K2能够促进BM-MSCs的成骨分化,并且可能通过成骨信号轴BMP-2/Smad/Runx2/Osterix来调控BM-MSCs的成骨分化过程。
英文摘要:
      Objective To study the influence and mechanism of Vitamin K2 on the osteogenic differentiation ability of bone marrow mesenchymal stem cells (BM-MSCs).Methods CCK-8 live cell assay was used to detect the influence of different concentrations of Vitamin K2 (1 nmol/L, 10 nmol/L, 100 nmol/L, 1 μmol/L, 10 μmol/L) at different times (24, 48, 72 h) on the growth of Bone Marrow Mesenchymal Stem Cells(BM-MSCs). QPCR was used to detect the expression level of bone morphogenetic protein (BMP-2) in the different concentrations of vitamin K2. Osteogenic signaling axis Smad1/5/8, Runx2 (Runt-related transcription factor-2), Osterix protein expression was detected by western blot after Vitamin K2 treatment. In order to further explore whether vitamin K2 regulates the osteogenic differentiation of BM-MSCs through the osteogenic signaling pathways Smad1/5/8, Runx2 and Osterix.Results Compared with the control group, low concentration (1 nmol/L) of vitamin K2 did not affect the growth activity of BM-MSCs, and medium concentration (10 nmol/L, 100 nmol/L, 1 μmol/L) significantly promoted the growth of BM-MSCs , High concentration (10 μmol/L) significantly inhibited cell growth (P<0.05). The medium concentration of vitamin K2 dose-dependently promoted the expression of BMP-2 mRNA and the formation of calcium nodules during the osteogenesis of BM-MSCs, and 1μmol /L vitamin K2 significantly promoted the expression levels of smad1/5/8, Runx2 and Osterix protein.ConclusionVitamin K2 could promote the osteogenic differentiation of MSCs, and it might regulate the osteogenic differentiation process of BM-MSCs through the osteogenic signal axis BMP-2/Smad/Runx2/Osterix.
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