A20调控NF-κB对强直性脊柱炎小鼠模型韧带骨化作用机制研究

The NF-κB signal pathway which regulate immune inflammatory response is one of the key point on inducing vertebral ligament ossification in ankylosing spondylitis,it hopefully make a breakthrough of the treatment in ankylosis with AS patients via suppressing NF-κB signal way.Recent study indicated that the TNF-induced protein 3(A20, TNFIAP3) is an inner inhibitor of NF-κB signal activation.However,the mechanism and function is not explicit. Our research plan to carry out the following points: ①Analysing the function of A20 gene and NF-κB signal pathway in ligament ossification of ankylosing spondylitis from hereditism and proteomics, exploring the relationship between multi-cytokines of NF-κB signal and A20 gene. ②Exploring the mechanism on A20 regulating inflammatory response through the activation of NF-κB signal pathway,blocking the unbalanced homeostasis of fibroblast function which transform to primitive osteoblast inducing the proliferation and mature.Our research strive to demonstrate the mechanism of A20 on regulating the murine AS model of ligament ossification,aiming to searching the new therapy for ankylosis of AS.

NF-κB作为调控炎性免疫应答的信号通路，在强直性脊柱炎（AS）椎体韧带骨化发挥了关键性调控作用，抑制该通路的活化有望治疗AS关节强直。新近研究发现TNF诱导蛋白3（A20，TNFIAP3）是一种NF-κB信号活化的内源性抑制剂，但作用和机制不明。本研究拟开展：①从基因和蛋白层面分析A20基因和NF-κB通路对AS小鼠模型韧带骨化中的作用，深入研究它与NF-κB多种活化信号分子的关系。②探讨在模拟AS病理成骨环境下，A20如何通过调控NF-κB信号通路活性，从而抑制炎症反应，阻断稳态失衡的成纤维细胞向原始成骨细胞分化、成熟及相关功能的影响。本研究力图阐明A20基因和蛋白对AS小鼠模型韧带骨化的功能和作用机制，期望为AS关节强直的治疗提供新的思路。

《A20调控NF-κB对强直性脊柱炎小鼠模型韧带骨化作用机制研究》的主要研究内容有如下四个部分，一为观察A20基因在AS模型小鼠的调控机制，如何阻断FLS-OB分化，A20在韧带骨化组表达下降，所得结果为后续实验的指导存在着重大的意义；第二部分为，探索A20基因通过抑制TNF-a和BMP-2、bFGF及Cbfal介导阻断病理性成骨细胞生成，结果发现A20治疗组可增加Treg数目，有效抑制炎症反应，该部分内容意在明确A20在强直性脊柱炎发病过程中所发挥的保护性抑或破坏性作用及其地位；第三部分在于观察A20在体内抑制NF-KB信号活性并且维持成纤维细胞分化稳态，结果发现A20抑制成纤维细胞骨化，但TGF-Beta有所升高。第四部分主要研究探索A20基因在韧带骨化中抑制病理成骨作用及其机制，以初步阐明A20调控韧带骨化的可能机制，结果发现A20治疗组可显著抑制MAPK通路及NF-kB的活化，从而抑制炎症发生，并在韧带骨化中发挥保护作用。该课题按实验计划进展顺利，为A20在治疗强直性脊柱炎的韧带骨化探索打下了基础。