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背景:力学信号与骨骼系统的生长发育、修复重建和疾病发生发展有密切联系,其对骨髓间充质干细胞的影响和作用机制值得关注。目的:探讨静水压力刺激对骨髓间充质干细胞分化的影响及相关机制。方法:①短期实验:将人骨髓间充质干细胞分别接种于正常DMEM培养基、成骨诱导培养基或含细胞外信号调节激酶1/2抑制剂U0126的DMEM培养基中,利用自制压力加载系统对其施加0,40,80 kPa的静水压强1,4 h。②长期实验:将人骨髓间充质干细胞分别接种于正常DMEM培养基或成骨诱导培养基,施加40 kPa的静水压强4 h/d,持续14 d,以不施加静水压的细胞作对照。结果与结论:实时定量反转录PCR结果显示,经成骨诱导或40 kPa静水压刺激4 h后,骨髓间充质干细胞内核心结合因子α1、骨钙素mRNA表达增加,过氧化物酶体增殖物活化受体γ2和脂肪酶mRNA表达降低,80 kPa静水压刺激没有出现这一规律;40 kPa静水压的成骨诱导作用可被U0126部分拮抗。组织化学染色显示40 kPa静水压刺激7 d,骨髓间充质干细胞碱性磷酸酶表达和活性均增加;持续刺激14 d,过氧化物酶体增殖物活化受体γ2和脂肪酶mRNA表达增加。说明一定强度和作用时间的静水压刺激可调节骨髓间充质干细胞分化,其作用部分依赖于细胞外信号调节激酶1/2信号途径。
BACKGROUND: Mechanical signals are closely related to the growth and development, repair and reconstruction of skeletal systems and the development of diseases. Their effects on bone marrow mesenchymal stem cells and their mechanisms of action are worthy of attention. Objective: To investigate the effect of hydrostatic pressure on the differentiation of bone marrow mesenchymal stem cells and its related mechanisms. Methods: ① Short-term experiment: human bone marrow-derived mesenchymal stem cells were inoculated into normal DMEM medium, osteogenic medium or DMEM medium containing extracellular signal-regulated kinase 1/2 inhibitor U0126, respectively, using self-made pressure loading system The hydrostatic pressure at 0, 40, 80 kPa was applied for 1,4 h. ② Long-term experiment: Human bone marrow-derived mesenchymal stem cells were inoculated into normal DMEM medium or osteogenic medium respectively and hydrostatic pressure of 40 kPa was applied for 4 h / d for 14 days. . RESULTS AND CONCLUSION: Real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) results showed that after induced by osteogenic induction or hydrostatic pressure of 40 kPa for 4 h, the expression of core-associated factor α1 and osteocalcin mRNA increased in bone marrow mesenchymal stem cells. Peroxidase The expression of activated receptor γ2 and lipase mRNA decreased, while the hydrostatic pressure at 80 kPa did not show this regulation. The osteoinductive effect of hydrostatic pressure at 40 kPa could be partially antagonized by U0126. Histochemical staining showed that the expression and activity of alkaline phosphatase in bone marrow mesenchymal stem cells increased after hydrostatic pressure of 40 kPa for 7 days. The mRNA expression of peroxisome proliferator-activated receptor γ2 and lipase increased . It indicates that the hydrostatic pressure stimulation with certain intensity and action time can regulate the differentiation of BMSCs, and its effect depends partly on the extracellular signal-regulated kinase 1/2 signaling pathway.