体外震波干预糖皮质激素诱导人股骨头BMECs损伤的11β-HSD基因表达研究

Bone microvascular endothelial cells (BMECs) damage is the fundamental pathological change of glucocorticoid(GC)-induced osteonecrosis(ON). Our previous study found that extracorporeal shock waves (SW) can reverse GC-induced human femoral head vascular endothelial injury. p38MAPK has been proved to be involved in mediating endothelial metabolic disorders. 11 beta-hydroxysteroid dehydrogenase (11β-HSD) is the GC-prereceptor modulator at the local tissue level, which can regulate the local level of the active hormone. We hypothesize that extracorporeal SW can down-regulate the expression of 11β-HSD gene and alleviate the GC-induced damage of human femoral BMECs mediated by the gene. Based on the three-dimensional model of GC-induced BMECs damage built by our research team, this study will explore the effect of GC on the expression of 11β-HSD in the BMECs of the femoral head measured by RT-PCR technique, observe the gene function of 11β-HSD via the overexpression and knocking down tests and reveal the dynamic changes of the above molecular biological index after SW protective intervention in 11β-HSD of BMECs injury in many aspects, then clarify the correlation between p38MAPK signaling pathway and shock wave process. We try to ascertain the possibility of potential clinical targeted therapy in bone necrosis so as to prevent bone necrosis in super early stage from the source.

骨内微血管内皮细胞(BMECs)损伤是糖皮质激素(GC)诱导股骨头坏死主要的发病机制。我们前期研究发现，体外震波可拮抗GC诱导人股骨头血管内皮损伤。p38MAPK信号通路被证明参与介导内皮功能代谢紊乱。GC受体前调节剂11β-羟基类固醇脱氢酶(11β-HSD)能调控局部组织激素活化水平。我们假设体外震波能下调11β-HSD基因表达，减轻其介导的激素性人股骨头BMECs损伤。本项目拟基于课题组前期建立的激素性人股骨头BMECs损伤三维模型，检测BMECs损伤11β-HSD 基因表达变化，通过过表达与基因沉默实验观察11β-HSD 基因的功能作用，探讨震波对激素性人股骨头BMECs损伤11β-HSD 基因的干预效果及作用机制，阐明p38MAPK在震波干预过程中的作用。构建骨坏死动物模型加以验证。揭示体外震波防治骨坏死的机制，有望实现激素受体前超早期干预，从源头上减少或避免激素性骨坏死的发生。

骨内微血管内皮细胞(BMECs)损伤是糖皮质激素(GC)诱导股骨头坏死主要的发病机制。GC受体前调节剂11β-羟基类固醇脱氢酶(11β-HSD)能调控局部组织激素活化水平。本研究建立糖皮质激素诱导人股骨头BMECs损伤的三维模型，找到最佳的激素干预浓度。免疫组织化学染色检查发现人股骨头内富含H亚型血管内皮细胞。利用RT-PCR技术和Western-blot方法检测糖皮质激素诱导人股骨头不同区域BMECs损伤的11β-HSD 基因表达变化差异，初步探明了人股骨头内微血管内皮细胞11β-HSD 与激素的作用关系，阐述了激素性股骨头BMECs 损伤的受体前调控机制。探究体外震波对激素性人股骨头内微血管内皮细胞损伤的干预效果，构建股骨头坏死动物模型进行了初步的验证。有助于进一步探索股骨头坏死靶向治疗可能的作用靶点，阐释了体外震波防治骨坏死的机制及应用前景，促使股骨头坏死GC 受体前超早期干预成为可能，具有较大的潜在应用价值。