老年黄韧带骨化症软骨细胞miR24成骨相关的分子网络及机制探究

项目名称： 老年黄韧带骨化症软骨细胞miR24成骨相关的分子网络及机制探究

项目编号： No.81471403

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 雪原

作者单位： 天津医科大学

项目金额： 80万元

中文摘要： 老年黄韧带骨化（ossification ligamentum flavum, OLF）症是非瘤性老年成骨现象，探索其成骨机制有利于防治老年OLF和骨质疏松（osteoporosis,OP）症。前期工作发现，老年OLF软骨细胞miR24是对照组表达量的1/2，并证实miR24抑制成骨伴有Tcf1降低。但目前尚不明确miR24成骨功效的确切转录后调控网络，尤其是αv整合素黏附（3D黏附）状态下的成骨调控网路，缺乏miR24抑制OLF和预防OP的证据。本次申报：1、检测αv整合素黏附状态和常规培养状态下，miR24对全转录组测序影响,分析与β-catenin/Tcf1相互作用的细胞内通路； 2、在体外筛选和验证miR24/β-catenin/Tcf1相关分子网络；3、分别建立OLF和OP小鼠的miR24转染动物模型，在体内验证miR24升高对OLF和miR24降低对OP的防治作用。

中文关键词： 微小RNA；黄韧带骨化症；软骨细胞；成骨过程；分子机制

英文摘要： MicroRNAs (miRNAs) are critical posttranscriptional regulators of gene expression that control osteoblast mediated bone formation. Deregulation of miRNA mediated mechanisms is emerging as an important pathological factor in bone degeneration (eg, osteoporosis) and other bone-related diseases. MiRNAs are intriguing regulatory molecules that are networked with cell signaling pathways and intricate transcriptional programs through ingenuous circuits with remarkably simple logic. Ossification of the ligamentum flavum (OLF) is one of the important causes of the stenosis of the spinal canal. OLF can result in slowly progressive paraphrases or acute paraplegia. OLF usually exists with myelopathy in the setting of thoracic spinal, but its etiology is not well defined. Our previous study first showed that the miR-24 was expression as half level of the normal ligament flavum, and the miR-24 down regulation could promote the bone formation with the Tcf1 level increase. But how or which molecular networks are involved the miR-24 regulating the post-transcription bone formation have not definited yet. The present study is aimed at illustration which nolecular netword was engaged in the bone formation under the miR-24 down-regulation. 1) the miR-24 is constructed expression and knocked in the MCSc，OLF chondrocytes and the ligamtum flavum fibroblast, the target genes are analysis by the slice. Bone formation is also evaluated. 2) the above step is repeated under the 3D matrix cultured 3) in vivo function of miR-4 in bone formation is exam by the cre-loxp mice, the miR-24 potential usage in the OLF and osteoporosis treatment is evaluated.

英文关键词： miRNA;ossification of ligamentum flavum;chondrocytes;bone formation process;molecular mechanism