以TNF信号通路为靶目标治疗类风湿关节炎的基础研究

TNF is the major cytokine driving inflammation in rheumatoid arthritis (RA) which characterized by synovial inflammation and joint destruction,TNF induces bone loss in common bone diseases by promoting osteoclast formation directly and indirectly.Developing new drug with anti-resorptive or anabolic effect, particularly, with dual anti-resorptive and anabolic effect simultaneously, offer a great promise for the prevention and treatment of osteoporosis and synovial inflammation. We found that TNF receptor associated factor 3 (TRAF3) plays important dual role to inhibit osteoclast and also inhibt synovial inflammation differentiation by negatively regulating NF-kB inducing kinase (NIK) that controls the activation of non-canonical NF-kB signaling pathway. In this proposal, we will investigate (1)TNF switches the differentiation of M-CSF-induced Ly6C-Gr1- M2 to Ly6C+Gr1-CD11c＋ and Ly6C-Gr1-CD11c＋inflammatory M1 macrophages.（2)Biphasic effect of RelB on OC formation and synovial inflammation .(3)if NIK inhibitor can prevent ovariectomy-induced osteoporosis in mice by inhibiting bone resorption , synovial inflammation and also stimulating bone formation.Completion of the proposed research not only determine the novel molecular mechanism of rheumatoid arthritis (RA),but also provide a lead compound that therapy of rheumatoid arthritis (RA).

类风湿关节炎是一种慢性关节疾病，慢性滑膜炎和关节损伤是其主要表现，TNF在类风湿关节炎中是主要的致炎因子，TNF可以直接或间接地促进破骨细胞形成而引起骨量流失。开发抗骨吸收或促进骨合成，抑制滑膜炎症的药物，对类风湿关节炎的防治有远大前景。实验发现TNF受体相关因子3（TRAF3）通过负性调控调节NF-kB诱导NIK所控制的非经典NF-kB通路而抑制骨吸收，同时抑制滑膜损伤。项目将探讨（1）TNF转化M-CSF诱导的Ly6C-Gr1- M2至Ly6C+Gr1-CD11c＋和Ly6C-Gr1-CD11c＋ 炎性的M1巨噬细胞。（2）NF-kB前体蛋白RelB在影响破骨细胞形成和滑膜增生的双相作用。（3）NIK抑制剂是否可以阻断非经典NF-kB通路而抑制破骨细胞和滑膜增生、促进成骨细胞分化。本项目完成不仅可以了解类风湿关节炎（RA）的分子机制，也可以对获得治疗类风湿关节炎的先导化合物有指导作用。

我们探讨了BMP-9在A549和NCI-H1650细胞增殖中的作用及其可能的分子机制。从招募的25名非小细胞肺癌（NSCLC）患者来评估BMP-9的mRNA表达，以确定其临床病理学意义。我们发现，重组蛋白BMP-9和BMP-9的过度表达促进A549和NCI-H1650细胞在体外增殖，这是由磷脂蛋白酶3激酶（PI3K）抑制剂（LY294002）消除的。蛋白印迹实验结果显示，BMP-9显著激活了PI3K/Akt和Smad1/5信号通路。体外实验，BMP-9促进肿瘤生长和PI3K/Akt和Smad1/5信号通路在A549或NCI-H1650细胞系衍生异种移植模型中。BMP-9或BMP-9受体ALK1抑制A549细胞在体外和体内生长，这与调节PI3K/Akt和Smad1/5信号通路有关。这些结果表明，BMP-9通过PI3K/Akt和Smad1/5信号通路促进A549和NCI-H1650细胞增殖。