上海口腔医学

上海口腔医学 ›› 2021, Vol. 30 ›› Issue (6): 579-584.doi: 10.19439/j.sjos.2021.06.004

• 论著 • 上一篇    下一篇

SDF-1α/CXCR4信号轴介导柚皮素对骨髓间充质干细胞成骨分化的影响

白书林1,2, 王奕佩1,2, 金珂1,2, 罗小玲1,2, 徐晓梅1,2   

  1. 1.西南医科大学附属口腔医院 正畸科,四川 泸州 646000;
    2.西南医科大学 口颌面修复重建和再生实验室,四川 泸州 646000
  • 收稿日期:2020-08-27 修回日期:2021-03-04 发布日期:2022-03-09
  • 通讯作者: 徐晓梅,E-mail: xuxiaomei@hotmail.com
  • 作者简介:白书林(1994-),男,在读硕士研究生,E-mail:2383804292@qq.com
  • 基金资助:
    四川省医学会资助项目（S18002）; 泸州市科技局-西南医科大学联合项目（2020LZXNYDZ06）

Naringenin promotes osteogenic differentiation of BMSCs via SDF-1α/CXCR4 signaling axis

BAI Shu-lin1,2, WANG Yi-pei1,2, JIN Ke1,2, LUO Xiao-ling1,2, XU Xiao-mei1,2   

  1. 1. Department of Orthodontics, Affiliated Stomatology Hospital of Southwest Medical University. Luzhou 646000;
    2. Laboratory of Oral and Maxillofacial Reconstruction and Regeneration,Southwest Medical University. Luzhou 646000, Sichuan Province, China
  • Received:2020-08-27 Revised:2021-03-04 Published:2022-03-09

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摘要： 目的: 探讨柚皮素对骨髓间充质干细胞（bone mesenchymal stem cells,BMSCs）成骨分化的影响,SDF-1α/CXCR4信号轴是否参与介导柚皮素促进BMSCs成骨分化。方法: 分离、培养及鉴定大鼠BMSCs,CCK8法检测不同浓度柚皮素对BMSCs增殖的影响,检测碱性磷酸酶（alkaline phosphatase,ALP）活性;RT-qPCR法检测各组ALP、骨钙素（osteocalcin,OCN）、趋化因子受体4（CXCL receptor 4,CXCR4）和基质细胞衍生因子1α（stromal cell derived factor-1,SDF-1α）的表达,ELISA法检测各组CXCR4和SDF-1α蛋白的表达。采用SPSS 21.0软件包对数据进行统计学分析。结果: 细胞鉴定结果表明,培养细胞为高纯度的BMSCs。第1、3天,柚皮素对BMSCs增殖无显著影响（P>0.05）;第5天时,50 μg/mL柚皮素可促进BMSCs增殖;第7天时,不同浓度柚皮素均促进BMSCs增殖（P<0.05）;同时,ALP活性随时间推移逐渐增强。RT-qPCR结果表明,柚皮素组和成骨诱导液组中相关基因表达均较培养基组明显增加,而AMD3100组中各基因表达均受到显著抑制（P<0.05）。ELISA检测结果显示,各组CXCR4和SDF-1α蛋白表达随诱导时间增加均逐渐上调,且两者均在100 μg/mL柚皮素组表达最高。结论: 柚皮素可促进BMSCs增殖和成骨向分化,SDF-1α/CXCR4信号轴参与柚皮素对BMSCs的成骨分化,主要在BMSCs成骨分化早期阶段。

关键词: 骨髓间充质干细胞, 柚皮素, SDF-1α/CXCR4, 成骨分化

Abstract: PURPOSE: To explore the influence of naringenin on osteogenic differentiation of bone mesenchymal stem cells(BMSCs), and the role of SDF-1α/CXCR4 signaling axis in the osteogenic differentiation by naringenin. METHODS: BMSCs of the rats were isolated,cultured and tested. CCK-8 assay was used to explore the proliferation ability of BMSCs in different concentrations of naringenin, and alkaline phosphatase(ALP) activity was detected. RT-qPCR was used to detect the mRNA expression of ALP, OCN, CXCR4 and SDF-1α in different groups. The expressions of CXCR4 and SDF-1α protein in BMSCs during osteogenic differentiation in different experimental groups were detected by ELISA. SPSS 21.0 software package was used for statistical analysis of the data. RESULTS: The results of cell identification showed that the cultured cells were BMSCs. At 1 d and 3 d, all concentrations of naringenin had no significant effect on the proliferation of BMSCs; and at 5 d, 50 μg/mL of naringenin promoted proliferation of BMSCs;furthermore, at 7 d, all concentrations of naringenin promoted proliferation of BMSCs(P<0.05). ALP activity value gradually increased in each concentration over time. From the RT-qPCR experiment, the mRNA expression of ALP, OCN, CXCR4 and SDF-1α in the naringenin group and the osteogenic induction group was significantly increased compared with the medium group(P<0.05). ELISA assay showed that the protein expressions of CXCR4 and SDF-1α increased gradually in the four groups as time went on and the expression of two proteins was the highest in 100 μg/mL naringenin group. CONCLUSIONS: Naringenin can promote the proliferation and osteogenic differentiation of BMSCs. SDF-1α/CXCR4 signaling axis is involved in the osteogenic differentiation of BMSCs by naringenin,particularly in the early stage of BMSCs osteogenic differentiation.

Key words: BMSCs, Naringenin, SDF-1α/CXCR4, Bone regeneration

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