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| 压力通过FAK-PI3K-AKT信号通路影响硬脑膜细胞成骨分子机制研究 |
| Study on the molecular mechanism of pressure affecting the osteogenic process of dura mater cells through the FAK-PI3K-AKT signaling pathway |
| 投稿时间：2025-03-03 修订日期：2025-03-10 |
| DOI： |
| 中文关键词: 关键词：硬脑膜细胞 成骨 成骨诱导 压力 FAK通路 |
| 英文关键词: Keywords: Dural cells:Osteogenesis Osteogenesis induction Pressure FAK pathway |
| 基金项目:国家自然科学基金项目（面上项目，重点项目，重大项目） |
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| 中文摘要: |
| 摘要 目的：探讨压力刺激对硬脑膜细胞成骨分化的影响及其与FAK-PI3K-AKT信号通路的调控关系。方法：采用成骨诱导培养基处理硬脑膜细胞，通过倒置显微镜动态观察细胞形态学改变，应用茜素红染色法和碱性磷酸酶（ALP）染色评估钙结节形成情况，采用RT-PCR技术检测骨钙素（OCN）和骨桥蛋白（OPN）mRNA表达水平，并利用Western blot分析压力干预下FAK-PI3K-AKT信号通路关键蛋白的时序性表达特征。结果：经过成骨诱导处理后，硬脑膜细胞的形态学特征发生了显著变化，由原本的梭状逐渐转变为短梭形或多边形，细胞体积显著增大，胞体变得细长且扁平，细胞核的体积也明显扩张。茜素红染色和碱性磷酸酶染色结果显示，成骨诱导后的染色程度相较于诱导前有了明显的加深。在压力刺激的作用下，硬脑膜细胞的FAK-PI3K-AKT信号通路表现出时间依赖性的激活特性，其表达水平随着时间的延长而逐渐上升。在诱导和压力后碱性磷酸酶（ALP）和骨钙素（OCN）等成骨蛋白的表达水平均显著上调。结论：硬脑膜细胞在成骨诱导后，通过茜素红和碱性磷酸酶染色观察到成骨结节随时间增多。压力对硬脑膜细胞的FAK/P13K/AKT信号通路蛋白表达及PCR检测的成骨相关基因表达均升高。 |
| 英文摘要: |
| Abstract Objective: To investigate the effects of mechanical stress on osteogenic differentiation of dural cells and its regulatory relationship with the FAK-PI3K-AKT signaling pathway.
Methods: Dural cells were treated with osteogenic induction medium. Morphological changes were dynamically observed under an inverted microscope. Alizarin red staining and alkaline phosphatase (ALP) staining were employed to evaluate calcium nodule formation. qRT-PCR was used to detect mRNA expression levels of osteocalcin (OCN) and osteopontin (OPN), while Western blot was performed to analyze the time-dependent expression characteristics of key proteins in the FAK-PI3K-AKT signaling pathway under mechanical stress.Results: Following osteogenic induction, dural cells exhibited significant morphological alterations: spindle-shaped cells transformed into short fusiform or polygonal forms, with enlarged cell bodies, elongated flattened morphology, and markedly increased nuclear volume. Alizarin red and ALP staining demonstrated intensified calcified nodule formation post-induction. Mechanical stimulation induced time-dependent activation of the FAK-PI3K-AKT pathway, with protein expression levels progressively increasing over time. Concurrently, osteogenic markers including ALP and OCN were significantly upregulated.Conclusion: Osteogenic induction promotes calcified nodule formation in dural cells, as evidenced by alizarin red and ALP staining. Mechanical stress enhances both the expression of FAK/PI3K/AKT pathway proteins and osteogenesis-related genes, suggesting a critical regulatory role of this pathway in stress-mediated osteogenic differentiation of dural cells. |
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