中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (31): 8047-8053.doi: 10.12307/2026.849

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

低氧预处理骨髓间充质干细胞旁分泌的细胞因子促进人脐静脉内皮细胞血管生成

刘  雷，毛  文，祁  福，李  红，张明焕   

  1. 武汉市第三医院关节创伤骨科，湖北省武汉市  430071
  • 收稿日期:2025-10-29 接受日期:2026-01-15 出版日期:2026-11-08 发布日期:2026-05-21
  • 通讯作者: 张明焕，副主任医师，武汉市第三医院关节创伤骨科，湖北省武汉市 430071
  • 作者简介:刘雷，男，1989年生，2016年武汉大学毕业，硕士，主治医师，主要从事创伤骨科相关研究。
  • 基金资助:
    武汉市科技局2022年度知识创新专项曙光计划项目(2022020801020550)，项目负责人：刘雷

Hypoxia preconditioning of bone marrow mesenchymal stem cells promotes angiogenesis in human umbilical vein endothelial cells through paracrine cytokines

Liu Lei, Mao Wen, Qi Fu, Li Hong, Zhang Minghuan   

  1. Department of Orthopedics for Joint Trauma, Wuhan Third Hospital, Wuhan 430071, Hubei Province, China
  • Received:2025-10-29 Accepted:2026-01-15 Online:2026-11-08 Published:2026-05-21
  • Contact: Zhang Minghuan, Associate chief physician, Department of Orthopedics for Joint Trauma, Wuhan Third Hospital, Wuhan 430071, Hubei Province, China
  • About author:Liu Lei, MS, Attending physician, Department of Orthopedics for Joint Trauma, Wuhan Third Hospital, Wuhan 430071, Hubei Province, China
  • Supported by:
    2022 Knowledge Innovation Special Program-Shuguang Project of Wuhan Municipal Science and Technology Bureau, No. 2022020801020550 (to LL)

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摘要：

文题释义：

骨髓间充质干细胞：是从大鼠或人骨髓中分离提取的一类成体干细胞，具有贴壁生长和多向分化潜能等特性。此研究的核心在于对骨髓间充质干细胞进行低氧预处理，以模拟体内组织缺血微环境，从而量化地激活旁分泌功能，使骨髓间充质干细胞分泌的促血管生成细胞因子水平显著提升。因此，文中的骨髓间充质干细胞是旁分泌活性因子工厂，经过低氧预处理后分泌的细胞因子是影响损伤内皮细胞功能的关键变量。
血管生成：此研究中的“血管生成”特指在细胞水平上，由损伤的内皮细胞所执行的一个体外量化过程，它是组织修复和再生的关键环节。文章关注的并非生物体完整的血管新生，而是核心步骤的体外模拟：即损伤后的内皮细胞在骨髓间充质干细胞条件培养基的刺激下，发生迁移、增殖并最终连接形成管腔样结构的能力。因此，“血管生成”在此处是一个可测量的、由细胞因子驱动的高阶功能学结果，直接反映了治疗策略的有效性。

摘要
背景：借鉴体内缺血缺氧微环境的特点，低氧预处理作为一种能模拟生理病理条件的主动策略，可显著增强干细胞的旁分泌功能，但低氧预处理对损伤内皮细胞血管生成行为的特异性影响与深层机制尚待深入解析。
目的：探讨低氧预处理对骨髓间充质干细胞旁分泌功能的调控作用，旨在定量探讨低氧预处理骨髓间充质干细胞条件培养基对损伤内皮细胞成血管能力的影响及机制。
方法：将人脐静脉内皮细胞分成对照组、模型组、常氧组和低氧组，对照组正常培养人脐静脉内皮细胞；模型组用1 μg/mL脂多糖处理24 h；常氧组和低氧组分别用常氧和低氧条件下培养48 h的骨髓间充质干细胞上清干预人脐静脉内皮细胞1 h，再用1 μg/mL脂多糖处理24 h。CCK-8检测细胞增殖能力，流式细胞术检测细胞凋亡率，小管形成实验检测细胞血管生成能力，ELISA检测细胞上清中肿瘤坏死因子α和白细胞介素6水平，RT-qPCR法和Western blot法检测细胞中细胞间黏附分子1、血管细胞黏附因子1和血管内皮生长因子 mRNA和蛋白的表达。
结果与结论：①与对照组相比，模型组细胞存活率、成管能力、细胞中细胞间黏附分子1、血管细胞黏附因子1和血管内皮生长因子 mRNA和蛋白表达水平显著降低(P < 0.01)，细胞凋亡率、细胞上清中肿瘤坏死因子α和白细胞介素6水平显著升高(P < 0.01)；②与模型组比较，常氧组上述指标差异均不明显(P > 0.05)，而低氧组细胞存活率、成管能力、细胞中细胞间黏附分子1、血管细胞黏附因子1和血管内皮生长因子 mRNA和蛋白表达水平显著升高(P < 0.01)，细胞凋亡率、细胞上清中肿瘤坏死因子α和白细胞介素6水平显著降低(P < 0.01)。结果表明，低氧预处理的骨髓间充质干细胞旁分泌细胞因子能够抑制血管内皮细胞的炎症反应，并通过上调血管生成因子促进血管生成。

关键词: 骨折愈合, 低氧预处理, 骨髓间充质干细胞, 血管生成, 旁分泌细胞因子, 炎症 

Abstract: BACKGROUND: Drawing on the characteristics of the in vivo ischemic and hypoxic microenvironment, hypoxic preconditioning, as an active strategy that simulates physiological and pathological conditions, can significantly enhance the paracrine function of stem cells. However, the specific effects and underlying mechanisms of hypoxic preconditioning on the angiogenic behavior of damaged endothelial cells remain to be further elucidated.
OBJECTIVE: To investigate the regulatory effect of hypoxic preconditioning on the paracrine function of bone marrow mesenchymal stem cells, aiming to quantitatively explore the effects and mechanisms of conditioned medium from hypoxic preconditioned bone marrow mesenchymal stem cells on the angiogenic capacity of damaged endothelial cells.
METHODS: Human umbilical vein endothelial cells were divided into control group, model group, normoxic group, and hypoxic group. The control group consisted of human umbilical vein endothelial cells cultured under normal conditions. The model group was treated with 1 μg/mL lipopolysaccharide for 24 hours. The normoxic and hypoxic groups were treated with supernatant from bone marrow mesenchymal stem cells cultured under normoxic and hypoxic conditions for 48 hours, respectively, for 1 hour, followed by treatment with 1 μg/mL lipopolysaccharide for 24 hours. Cell proliferation was detected by CCK-8 assay. Cell apoptosis rate was determined by flow cytometry. Cell angiogenesis capacity was measured by tube formation assay. The levels of tumor necrosis factor-α and interleukin-6 in the cell supernatant were detected by ELISA. The mRNA and protein expression levels of intercellular adhesion molecule 1, vascular cell adhesion molecule 1, and vascular endothelial growth factor in cells were detected by RT-qPCR and western blot assay, respectively. 
RESULTS AND CONCLUSION: (1) Compared with the control group, the model group showed significantly lower cell survival rate, tube formation ability, and mRNA and protein expression levels of intercellular adhesion molecule 1, vascular cell adhesion molecule 1, and vascular endothelial growth factor (P < 0.01), while the apoptosis rate and levels of tumor necrosis factor α and interleukin 6 in the cell supernatant were significantly higher (P < 0.01). (2) Compared with the model group, there were no significant differences in the above indicators in the normoxic group (P > 0.05), while the hypoxic group showed significantly higher cell survival rate, tube formation ability, and mRNA and protein expression levels of intercellular adhesion molecule 1, vascular cell adhesion molecule 1, and vascular endothelial growth factor (P < 0.01), and significantly lower apoptosis rate and levels of tumor necrosis factor α and interleukin 6 in the cell supernatant (P < 0.01). The results indicate that paracrine cytokines from hypoxic preconditioned bone marrow mesenchymal stem cells can inhibit the inflammatory response of vascular endothelial cells and promote angiogenesis by upregulating angiogenic factors. 

Key words: fracture healing, hypoxia pretreatment, bone marrow mesenchymal stem cells, angiogenesis, paracrine cytokines, inflammation

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