Ihh信号通路在周期性机械应力刺激诱导黄韧带骨化中的作用机制

项目名称： Ihh信号通路在周期性机械应力刺激诱导黄韧带骨化中的作用机制

项目编号： No.81501917

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 高瑞

作者单位： 中国人民解放军第二军医大学

项目金额： 18万元

中文摘要： 黄韧带骨化是一种能引起脊髓严重受压的致瘫性疾病，其发生和进展与脊柱失稳引起的局部应力增加密切相关，但具体分子机制不清。Ihh信号通路是骨生长发育的重要信号通路之一，通过浓度梯度来调控细胞行为。我们前期实验发现黄韧带骨化标本中Ihh的表达异常增高。对骨化的黄韧带细胞进行周期性机械应力刺激后，Ihh和骨化标记物的mRNA同时明显上调。抑制Ihh信号通路后，骨化标记物mRNA的上调受到抑制。我们推测：Ihh信号通路参与了周期性机械应力刺激诱导黄韧带骨化的过程。本研究拟在黄韧带骨化的病理组织水平检测Ihh分泌调控分子及Ihh信号通路相关蛋白的表达。在细胞水平分别沉默Ihh分泌调控分子及信号通路中关键蛋白的基因，明确周期性应力刺激诱导黄韧带骨化过程中Ihh的浓度梯度形成机制和信号传导机制。通过构建Ihh基因沉默小鼠的黄韧带骨化模型进一步验证上述机制。本研究将为黄韧带骨化的发病机制及治疗提供新的思路。

中文关键词： 黄韧带骨化；Ihh信号通路；周期性机械应力；分子机制

英文摘要： Ossification of ligamentum flavum (OLF) is one of the main causes of thoracic myelopathy. It is reported OLF is closely correlated with the increased local stress caused by the instability of the spine; however, the exact mechanism is unclear. Indian hedgehog (Ihh) signaling pathway is one of the important signaling pathways in bone development, which regulate cell differentiation by development of the concentration gradient. We have found significant overexpression of Ihh in OLF samples. Furthermore, application of cyclic mechanical stress to OLF cells resulted in significant increases in mRNA expression levels of Ihh and the ossification markers, while inhibition of Ihh resulted in the inhibited increase in mRNA expression levels of the ossification markers. Thus, we concluded that Ihh play an important role in OLF development. In this study, we will firstly examine the secretion regulatory elements of Ihh, and the expression of proteins of the Ihh pathway in OLF sample. Then, silence the genes of the secretion regulatory elements of Ihh and the genes of key proteins of the Ihh pathway, to confirm the mechanisms of development of the concentration gradient and the signal transduction of Ihh subjected to cyclic mechanical stress. Finally, establish an animal model of OLF induced by cyclic mechanical stress by using Ihh gene silence mouse, to test the above mechanisms. This study could provide a new clue to pathogenesis and treatment of OLF.

英文关键词： ossification of ligamentum flavum ;Indian hedgehog signaling pathway; cyclic mechanical stress ;molecular mechanism