miR-27a调控Galectin-3促进纤维增生在黄韧带肥厚中的作用及机制研究

Ligamentum flavum hypertrophy (LFH) is one of important causes of degenerative spinal stenosis. It has been confirmed that fibrosis is the main pathological basis of ligamentum flavum hypertrophy. However, the regulatory mechanism has not been clear. Our research found that miR-27a expression in hypertrophied ligamentum flavum had been down-regulated and the pilot experiment showed that Galectin-3 expression might depend on the target regulation of miR-27a. Meanwhile, the up-regulation of Galectin-3 played an important role in LFH. However, the mechanisms of Galectin-3 in regulation of fibrosis and miR-27a mediating Galectin-3 in LFH have needed to be explored and proven. Therefore, the research should first study the clinical data and ligamentum flavum specimens from patients. Then, the fibroblasts should be cultured from hypertrophied ligamentum flavum in this study to clarify the role of miR-27a in LFH and its regulatory role on Galectin-3. Finally, it should be validated through establishing a mouse model for LFH in vitro. This project should provide the experimental basis for clarify the role and mechanism of miR-27a and Galectin-3 in LFH. In addition, it is benefit to reveal new understandings of pathogenesis and new target genes for further gene therapy.

黄韧带肥厚是退变性椎管狭窄的重要原因之一。研究表明，纤维增生是黄韧带肥厚的主要病理基础，但其调控机制尚未明确。项目组研究发现miR-27a在肥厚黄韧带中表达显著下调，而且预实验结果提示其可能靶向调控半乳糖凝集素-3（Galectin-3）的表达，而后者表达升高可能促进了黄韧带肥厚的进展。但是，Galectin-3促进纤维增生的深入作用机制仍有待阐明，且miR-27a对此作用的调控也需要进一步证实。因此，本项目拟首先对患者临床资料及黄韧带标本进行分析；然后，体外培养黄韧带成纤维细胞，对miR-27a在黄韧带肥厚中的作用，及其调控Galectin-3表达的作用及机制进行深入研究；最后，通过建立黄韧带肥厚小鼠模型进行体内验证。本课题将为明确miR-27a调控Galectin-3表达在黄韧带肥厚中的作用及机制提供实验依据，有助于重新认识黄韧带肥厚的发病机理，为广大椎管狭窄症患者提供精准治疗新靶点。

项目组从患者临床资料和黄韧带标本进行分析，采用免疫组化染色、定量PCR和WB等方法比较肥厚与正常的黄韧带中多种分子的表达差异。然后，体外培养黄韧带成纤维细胞，对miR-27a-3p在抑制纤维增生的作用及其调控半乳糖凝集素-3（Galectin-3，Gal-3）表达的作用进行研究。最后，为了体内验证miR-27a-3p作用，建立了大鼠黄韧带肥厚模型。项目组研究发现miR-27a-3p在肥厚黄韧带中表达显著下调，这可能是黄韧带肥厚的重要因素之一。外源补充miR-27a-3p可以抑制纤维增生，这对改善黄韧带肥厚是有益的。