Department of Physiology, Guangxi Medical University, Nanning, People's Republic of China.
Key Laboratory of Longevity and Aging-Related Diseases, Ministry of Education, Guangxi Medical University, Nanning, People's Republic of China.
J Orthop Surg Res. 2023 May 30;18(1):391. doi: 10.1186/s13018-023-03859-6.
Osteoporosis is a common systemic bone disease that leads to bone fragility and increases the risk of fracture. However, the pathogenesis of osteoporosis is considered to be highly complex. The exosomes can regulate the communication between cells. The specific mechanism of information transmission between osteoblasts and endothelial cells is worthy of further study.
Exosomes were isolated and verified from senescent osteoblasts. The source and properties of exosomes were determined by TEM, particle size analysis and western blot. We established the co-culture model of endothelial cells and senescent osteoblasts. We used qRT-PCR to identify differentially expressed miRNAs. The functional changes of vascular endothelial cells were verified by cell transfection. β-Galactosidase cell senescence assay, Hoechst cell apoptosis assay, Ki67 cell proliferation assay and Transwell migration assay were used to verify cell senescence, apoptosis, proliferation, and migration. The potential target gene of miRNA was detected by bio-informatics pathway and double luciferase report.
We discovered that senescent osteoblasts could promote the senescence and apoptosis of vascular endothelial cells and inhibit their proliferation and migration. miR-214-3p was upregulated in senescent osteoblast-derived exosomes. miR-214-3p could effectively promote senescence and apoptosis of endothelial cells and inhibit proliferation and migration ability. L1CAM is a miR-214-3p direct target gene determined by bio-informatics and double luciferase report.
In conclusion, senescent osteoblast-derived exosomes can accelerate endothelial cell senescence through miR-214-3p/L1CAM pathway. Our experiments reveal the role of exosomes in the skeletal microenvironment.
骨质疏松症是一种常见的全身性骨骼疾病，会导致骨骼脆弱，增加骨折风险。然而，骨质疏松症的发病机制被认为非常复杂。外泌体可以调节细胞间的通讯。成骨细胞和内皮细胞之间信息传递的具体机制值得进一步研究。
从衰老的成骨细胞中分离和验证外泌体。通过 TEM、颗粒大小分析和 Western blot 确定外泌体的来源和特性。我们建立了内皮细胞和衰老成骨细胞的共培养模型。我们使用 qRT-PCR 鉴定差异表达的 miRNA。通过细胞转染验证血管内皮细胞的功能变化。β-半乳糖苷酶细胞衰老试验、Hoechst 细胞凋亡试验、Ki67 细胞增殖试验和 Transwell 迁移试验验证细胞衰老、凋亡、增殖和迁移。通过生物信息学途径和双荧光素酶报告检测 miRNA 的潜在靶基因。
我们发现衰老的成骨细胞可以促进血管内皮细胞的衰老和凋亡，抑制其增殖和迁移。衰老的成骨细胞来源的外泌体中 miR-214-3p 上调。miR-214-3p 可以有效促进内皮细胞的衰老和凋亡，抑制其增殖和迁移能力。L1CAM 是通过生物信息学和双荧光素酶报告确定的 miR-214-3p 的直接靶基因。
总之，衰老的成骨细胞衍生的外泌体可以通过 miR-214-3p/L1CAM 通路加速内皮细胞衰老。我们的实验揭示了外泌体在骨骼微环境中的作用。
