口腔医学研究 ›› 2022, Vol. 38 ›› Issue (2): 157-163.DOI: 10.13701/j.cnki.kqyxyj.2022.02.013

• 口腔干细胞研究 • 上一篇    下一篇

黄精多糖干预骨质疏松小鼠脂肪干细胞成骨分化的实验研究

陆诗1,2, 何清明1, 娄方芝2, 彭双麟2, 高俞锦1, 肖金刚1,2,3*   

  1. 1.西南医科大学口颌面修复重建和再生实验室 四川 泸州 646000;
    2.西南医科大学附属口腔医院口腔种植科 四川 泸州 646000;
    3.西南医科大学附属口腔医院口腔颌面外科 四川 泸州 646000
  • 收稿日期:2021-07-26 出版日期:2022-02-28 发布日期:2022-02-23
  • 通讯作者: *肖金刚,E-mail:drxiaojingang@163.com
  • 作者简介:陆诗(1996~ ),女,安徽阜阳人,硕士在读,主要从事口腔种植临床与基础研究。
  • 基金资助:
    国家自然科学基金(81870746);国家级大学生创新训练项目(201910632044,202010632001);四川省大学生创新训练项目(川教函2020[409]号);西南医科大学大学生创新训练项目(西南医大教[2019]14号、西南医大教[2020]5号)

Effect of Polygonatum Sibiricum Polysaccharides on Osteogenic Differentiation of Adipose Derived Stem Cells in Osteoporosis Mice

LU Shi1,2, HE Qingming1, LOU Fangzhi2, PENG Shuanglin2, GAO Yujin1, XIAO Jingang1,2,3*   

  1. 1. Luzhou Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Luzhou 646000, China;
    2. Department of Oral Implantology, the Affiliated Stomatology Hospital of Southwest Medical University, Luzhou 646000, China;
    3. Department of Oral and Maxillofacial Surgery, the Affiliated Stomatology Hospital of Southwest Medical University, Luzhou 646000, China
  • Received:2021-07-26 Online:2022-02-28 Published:2022-02-23

摘要: 目的: 研究不同浓度的黄精多糖(polygonatum sibiricum polysaccharides, PSP)对骨质疏松小鼠脂肪干细胞(adipose derived stem cells in osteoporosis mice, OP-ASCs)成骨分化的影响。方法: 建立骨质疏松症小鼠动物模型。体外分离培养OP-ASCs。使用不同浓度(5、10、25、50、75、100 mg/L)PSP干预OP-ASCs,成骨诱导后,CCK-8法检测细胞增殖;PSP干预OP-ASCs成骨诱导3 d和5 d后,Western blot及Real-time PCR检测成骨相关因子表达情况;碱性磷酸酶染色检测成骨能力改变。结果: PSP干预后第1、3、5、7天,相较于空白对照组,其他组OP-ASCs细胞增殖活力增加。其中PSP浓度为75 mg/L和100 mg/L时OP-ASCs细胞增殖活力较其他实验组有所下降(P<0.001)。在PSP浓度为50 mg/L时,Western blot及Real-time PCR结果显示:实验组Runx2、OPN、β-catenin、P-GSK-3β蛋白及其基因表达水平明显高于对照组(P<0.05);碱性磷酸酶染色示成骨效果实验组高于对照组。结论: PSP可以增强OP-ASCs成骨分化能力,可能是PSP通过上调Wnt/β-catenin信号通路实现。

关键词: 黄精多糖, 骨质疏松症, 脂肪干细胞, 成骨分化

Abstract: Objective: To study the effect of polygonatum sibiricum polysaccharides (PSP) on osteogenic differentiation of adipose derived stem cells in osteoporosis mice (OP-ASCs). Methods: Osteoporosis mouse model was established. OP-ASCs were isolated and cultured in vitro. Cell proliferation was detected by CCK-8 method after using different concentrations of PSP to intervene in OP-ASCs. Osteogenic capacity was analyzed by western-blot, real-time PCR, and ALP staining at day 3 and 5 after osteogenic differentiation in PSP-intervened OP-ASCs. Results: When the concentration of PSP was 5, 10, 25, and 50 mg/L, the proliferation of OP-ASCs was not significantly affected on day 3 and 5 (P>0.05). Western-blot and real-time PCR results showed that Runx2, OPN, β-catenin, P-GSK-3β protein and Runx2, Opn, β-catenin gene expression in the treatment group were significantly higher than those in the control group (P<0.05). ALP staining showed that the osteogenic effect was significantly higher than the control group. Conclusion: PSP can enhance osteogenic differentiation of OP-ASCs, which may be achieved by up-regulating Wnt/β-catenin signaling pathway.

Key words: polygonatum sibiricum polysaccharides, osteoporosis, adipose-derived stem cells, osteogenic differentiation