上海口腔医学

上海口腔医学 ›› 2022, Vol. 31 ›› Issue (1): 24-28.doi: 10.19439/j.sjos.2022.01.005

• 论著 • 上一篇    下一篇

雷奈酸锶对大鼠骨髓间充质干细胞成软骨分化的影响

俞灏1,2, 刘燕1,2, 杨翔文1,2, 张帆1,2, 何嘉菁1,2, 钟群1,2, 郭晓静1,2   

  1. 1.上海市口腔医院,上海 200001;
    2.上海市颅颌面发育与疾病重点实验室,上海 200001
  • 收稿日期:2020-08-27 修回日期:2020-11-15 出版日期:2022-02-25 发布日期:2022-03-10
  • 通讯作者: 郭晓静,E-mail：guo_xiaojing@fudan.edu.cn
  • 作者简介:俞灏（1988-）,男,硕士,主治医师,E-mail：yuhao_880514@fudan.edu.cn
  • 基金资助:
    上海市科委扬帆计划(19YF1442400); 上海市口腔病防治院院级课题(SSDC-2017-01、SSDC-2018-15)

Effect of strontium ranelate on chondrogenic differentiation of rat bone mesenchymal stem cells

YU Hao1,2, LIU Yan1,2, YANG Xiang-wen1,2, ZHANG Fan1,2, HE Jia-jing1,2, ZHONG Qun1,2, GUO Xiao-jing1,2   

  1. 1. Shanghai Stomatological Hospital & School of Stomatology, Fudan University. Shanghai 200001;
    2. Shanghai Key Laboratory of Craniomaxillofacial Development and Diseases, Fudan University. Shanghai 200001, China
  • Received:2020-08-27 Revised:2020-11-15 Online:2022-02-25 Published:2022-03-10

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摘要： 目的: 探讨雷奈酸锶（strontium ranelate,SrR）对大鼠骨髓间充质干细胞（bone mesenchymal stem cells,BMSCs）增殖及成软骨分化的作用。方法: 体外分离培养大鼠BMSCs,以含0.25~2.0 mmol/L的SrR软骨诱导分化培养基培养,CCK-8法检测细胞增殖状态,甲苯胺蓝及阿立新蓝染色观察成软骨分化情况,定量检测基质Hyp含量,PCR及Western免疫印迹（WB）定量或半定量检测相关基因及蛋白表达情况。采用SPSS 22.0软件包对数据进行统计学分析。结果: 0.25 mmol/L的 SrR不会抑制BMSCs增殖,基质硫酸软骨素及蛋白多糖含量较高。Hyp检测显示,0.25 mmol/L的SrR处理后细胞外基质胶原纤维含量较高。PCR及WB亦显示,0.25 mmol/L 的SrR处理细胞后Sox-9、Col-Ⅱ基因及蛋白和Aggrecan蛋白表达水平升高,MMP-9基因表达降低。结论: 0.25mmol/L 的SrR能显著促进大鼠BMSCs成软骨分化。

关键词: 雷奈酸锶, 骨髓间充质干细胞, 成软骨分化

Abstract: PURPOSE: The aim of present study was to explore the effect of strontium ranelate (SrR) on the proliferation and chondrogenic differentiation of rat bone mesenchymal stem cells (BMSCs). METHODS: Rat BMSCs were isolated and cultured in chondrogenic differentiation medium containing 0.25-2.0 mmol/L strontium ranelate. CCK-8 assay was used to study the influence of cell proliferation. Toluidine blue staining and alizarin blue staining were used to observe chondrogenic differentiation. Quantitative hydroxyproline (Hyp) activity assay was conducted. PCR and Western blots were used to detect the expression of related genes and proteins. Statistical analysis was performed using SPSS 22.0 software package. RESULTS: 0.25 mmol/L strontium ranelate did not inhibit the proliferation of BMSCs and promote the expression of chondroitin sulfate and proteoglycan. Hyp assay showed a higher content of collagen fibers in extracellular matrix in 0.25 mmol/L SrR treatment group. PCR and WB also showed up-regulated expression of Sox-9, Col-Ⅱ gene and protein, Aggrecan protein, and suppressed expression of MMP-9 gene. CONCLUSIONS: 0.25 mmol/L SrR could significantly promote chondrogenic differentiation of BMSCs.

Key words: Strontium Ranelate, Bone mesenchymal stem cells, Chondrogenic Differentiation

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