中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (35): 7481-7489.doi: 10.12307/2025.991

• 骨组织构建 bone tissue construction •    下一篇

川续断皂苷Ⅵ促进低氧环境下MC3T3-E1细胞的成骨分化

李蕴喆1，2，牛泽蕃1，2，王紫瑈2，艾崇毅2，陈  刚1，王新兴2   

  1. 1天津医科大学口腔医学院，天津市  300041；2军事科学院军事医学研究院，北京市  100000

  • 收稿日期:2024-11-18 接受日期:2024-12-31 出版日期:2025-12-18 发布日期:2025-04-30
  • 通讯作者: 陈刚，博士，教授，天津医科大学口腔医学院，天津市 300041 共同通讯作者：王新兴，博士，教授，军事科学院军事医学研究院，北京市 100000
  • 作者简介:李蕴喆，女，1996年生，山东省平度市人，汉族，天津医科大学在读硕士，主要从事骨再生相关研究。

Asperosaponin VI promotes osteogenic differentiation of MC3T3-E1 cells under hypoxia environment #br#
#br#

Li Yunzhe1, 2, Niu Zefan1, 2, Wang Zirou2, Ai Chongyi2, Chen Gang1, Wang Xinxing2   

  1. 1College of Stomatology, Tianjin Medical University, Tianjin 300041, China; 2Institute of Military Medical Research, Academy of Military Medical Sciences, Beijing 100000, China
  • Received:2024-11-18 Accepted:2024-12-31 Online:2025-12-18 Published:2025-04-30
  • Contact: Chen Gang, PhD, Professor, College of Stomatology, Tianjin Medical University, Tianjin 300041, China Co-corresponding author: Wang Xinxing, PhD, Professor, Institute of Military Medical Research, Academy of Military Medical Sciences, Beijing 100000, China
  • About author:Li Yunzhe, Master candidate, College of Stomatology, Tianjin Medical University, Tianjin 300041, China; Institute of Military Medical Research, Academy of Military Medical Sciences, Beijing 100000, China

PDF

77

摘要：


文题释义：
低氧：指空气中氧体积分数低于21%。当组织发生炎症、水肿时，由于毛细血管扩张和通透性增加，血液流速加快，氧气供应不足，局部细胞可能因缺氧变性、坏死，影响组织愈合。
川续断皂苷Ⅵ：又称木通皂苷D，是从中国传统中草药续断中提取的一种皂苷组分，是续断的主要活性成分，具有促进骨愈合、止血、抗老年痴呆、保护心脏、补肝肾等药理活性。

背景：川续断皂苷Ⅵ具有良好的促成骨作用，但其在低氧环境下促进细胞成骨的作用尚不明确。
目的：探讨川续断皂苷Ⅵ在低氧环境下对MC3T3-E1细胞成骨分化的影响及作用机制。
方法：将MC3T3-E1细胞分为3组，对照组细胞在常氧条件下(体积分数21%O2)用完全培养基培养，低氧组细胞在低氧条件下(体积分数0.5% O2)用完全培养基培养，川续断皂苷Ⅵ组细胞在低氧条件下(体积分数0.5%O2)用含有川续断皂苷Ⅵ的完全培养基培养。24 h后，采用CCK-8法和EdU染色法检测细胞增殖活性，TUNEL染色法和Western blot实验检测细胞凋亡情况，流式细胞术检测细胞内活性氧水平和细胞周期分布。另外，各组细胞在低氧条件下分别用含1×10-5，1×10-6，1×10-7 mol/L川续断皂苷Ⅵ的成骨诱导液培养7 d，通过光学显微镜观察和碱性磷酸酶染色筛选川续断皂苷Ⅵ最适浓度，采用Western blot方法检测骨形态发生蛋白2、骨桥蛋白、PI3K/AKT信号轴相关蛋白表达。
结果与结论：①与对照组相比，低氧条件下MC3T3-E1细胞增殖能力下降，1×10-6 mol/L川续断皂苷Ⅵ可以显著提高低氧状态下细胞增殖能力，并且减少细胞凋亡；②与低氧组相比，川续断皂苷Ⅵ组细胞内活性氧减少，进入S期细胞比例明显增多；③与低氧组相比，1×10-6 mol/L川续断皂苷Ⅵ组细胞形态舒展，较多突起，碱性磷酸酶染色较深；④与低氧组相比，川续断皂苷Ⅵ组骨形态发生蛋白2、骨桥蛋白表达增加，p-PI3K、p-AKT蛋白表达降低。结果表明，低氧环境下川续断皂苷Ⅵ可能通过调节PI3K/AKT通路促进MC3T3-E1细胞成骨分化。

https://orcid.org/0009-0003-1223-3282(李蕴喆)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: MC3T3-E1细胞, 低氧, 川续断皂苷Ⅵ, PI3K, AKT, 通路, 细胞分化, 成骨

Abstract: BACKGROUND: Asperosaponin VI has good osteogenic effects, but its ability to promote cellular osteogenesis under hypoxia environment is not yet clear.
OBJECTIVE: To investigate the effect and potential mechanism of Asperosaponin VI on osteogenic differentiation of MC3T3-E1 cells under hypoxia environment.
METHODS: MC3T3-E1 cells were divided into three groups. Cells in the control group were cultured in a complete medium under normoxic conditions (volume fraction of 21% O2); cells in the hypoxia group were cultured in the complete medium under hypoxia conditions (volume fraction of 0.5% O2); and cells in the Asperosaponin VI group were cultured in the complete medium containing Asperosaponin VI under hypoxia conditions (volume fraction of 0.5% oxygen). After 24 hours of culture, cell counting kit-8 method and EdU staining were used to detect cell proliferation activity, TUNEL staining and western blot assay were performed to detect cell apoptosis, and flow cytometry was used to detect intracellular reactive oxygen species levels and cell cycle distribution. Each group of cells was cultured in an osteogenic induction medium containing 1×10-5, 1×10-6, and 1×10-7 mol/L Asperosaponin VI under hypoxia conditions for 7 days. The optimal concentration of Asperosaponin VI for intervention was identified using alkaline phosphatase staining under optical microscopy. Western blot was used to detect the expression of bone morphogenetic protein 2, osteopontin, and PI3K/AKT signaling axis-related proteins.
RESULTS AND CONCLUSION: (1) Compared with the control group, the proliferation ability of MC3T3-E1 cells decreased under hypoxia conditions. 1×10-6 mol/L 
Asperosaponin VI could significantly improve the cell proliferation ability under hypoxia conditions and reduce cell apoptosis. (2) Compared with the hypoxia group, the Asperosaponin VI group showed a decrease in intracellular reactive oxygen species and a significant increase in the proportion of cells in the S phase. (3) Compared with the hypoxia group, the cell morphology in the 1×10-6 mol/L Asperosaponin VI group was elongated, with more protrusions and darker alkaline phosphatase staining. (4) Compared with the hypoxia group, the expression of bone morphogenetic protein 2 and osteopontin increased in the Asperosaponin VI group, while the expression of p-PI3K and p-AKT proteins decreased. These findings indicate that under hypoxia conditions, Asperosaponin VI can promote osteogenic differentiation of MC3T3-E1 cells, possibly by regulating the PI3K/AKT pathway.

Key words: MC3T3-E1 cell, hypoxia, Asperosaponin VI, PI3K, AKT, pathway, cell differentiation, osteogenesis

中图分类号: