体外震波作用于股骨头H亚型微血管内皮细胞miRNA干预激素性骨坏死形成的机制研究

Injury of bone microvascular endothelial cell is an important pathological mechanism for the development of glucocorticoid-induced osteonecrosis of the femoral head (ONFH); extracorporeal shockwaves, as a non-invasive mechanical stimulation, can effectively inhibit this injury. H subtype vessels couple bone formation and angiogenesis, and may play a major role in these processes. Funded by our former National Natural Science Foundation Project, by using the next generation sequencing and bioinformatic analysis, we selected 13 kinds microRNAs (miRNAs) , which were significantly upregulated or downregulated in bone microvascular endothelial cells (BMECs) after the intervention of extracorporeal shock wave (ESW). In order to investigate whether ESW acts a protective role by adjusting expression level of these miRNAs, we plan to construct three-dimensional BMECs model and mouse experimental model of glucocorticoid-induced ONFH and then by overexpressing or silencing these miRNAs, to explore their targeted molecules and downstream signal pathways in the development of glucocorticoid-induced ONFH. Then the targeted genes were further confirmed by the method of luciferase reporter gene, meanwhile, the change of angiogenesis and cancellous bone parameters was analyzed. The aim of this study is to reveal the molecular mechanism of extracorporeal shock wave intervention in the prevention of glucocorticoid-induced BMECs injury.

骨微血管内皮细胞(BMECs)损伤是激素性股骨头坏死的重要发病机制，体外震波作为一种无创的机械刺激，可有效干预这一损伤的发生，H亚型血管偶联了骨形成和血管形成，可能在其中起到关键作用。我们在前一个国自然项目的资助下，通过二代测序和生信分析筛选出13种miRNAs，其在体外震波干预后BMECs内表达量明显上调或下调。为进一步探索体外震波是否通过改变上述miRNAs的表达对H亚型血管起到保护作用，本研究拟构建激素性骨坏死的三维BMECs模型和小鼠动物实验模型，通过差异miRNA过表达/沉默的方式研究其在激素性BMECs损伤中的分子靶点和下游信号通路，通过双荧光素酶报告基因实验进一步验证其靶基因，同时观测分析体外震波作用于miRNA与血管改变和骨量改变的关系，进一步揭示体外震波干预激素性骨坏死的机制。

骨微血管内皮细胞(Bone microvascular cells，BMECs)损伤是激素性股骨头坏死的重要发病机制，体外震波(Extracorporeal shockwaves therapy, )作为一种无创的机械刺激，可有效干预这一损伤的发生。我们通过体内、体外实验发现激素可明显减少内皮细胞的表达，抑制其增殖并促进凋亡，并减低其迁移及成血管能力，而可一定程度的逆转激素其负面效应。我们进一步应用生物信息学对样本量为16例的BMECs差异表达芯片进行分析，发现7个差异，并根据表达差异显著程度选择-223-3p做进一步的靶基因预测及下游分子机制研究。通过转染人工合成的-223-3p inhibitor、过表达质粒、双荧光素酶报告基因实验等对的促增殖、促迁移、促血管生成及抗凋亡作用进行分子机制研究，进而阐明干预激素性股骨头坏死的分子生物学机制。