miR-150下调痛风炎症反应的靶点及作用机制研究

Gout, which is characterised by deposition of monosodium urate monohydrate (MSU) in synovial fluid and other tissues, is the most common form of inflammatory arthritis. Self-limiting is the pronounced feature of acute gout which different from other arthritis, while the mechanism is not clear. Increasing evidences demonstrated miRNAs can play as negtive regulator in infectious and autoimmune diseases. Preliminary results have confirmed that miR-150 participated in the MSU induced inflammation. In this study, we choice MyD88 (myeloid differentiation factor 88), P2X7R (the purinergic receptor P2X ligand-gated ion channel 7 receptor) and SOCS1 (suppressor of cytokine signaling 1) as the potential target of miR-150. Gain of function and loss of function will be used to detemine the real target and mechanism details. We will detect miR-150, the potential targets and related signaling molecules of miR-150, pro-inflammatory and anti-inflammatory level in different stages of gout development to reveal mechanism of miR-150 regulate seal-limiting of acute gout and the role of miR-150 in development of gout. The project will reveal the pathophysiology of gout in the aspect of miRNAs immune regulating gout and provide theoretical basis for clinical targeted therapy.

痛风是尿酸盐(monosodium urate, MSU)晶体沉积于关节及周围组织而引起的炎症反应，其区别于其他关节炎的一个显著特征为急性炎症具有自限性，但具体机制不详。miRNAs在多种炎症和自身免疫性疾病中能够起到负性调控作用。前期工作已经证实miR-150参与了MSU诱发的炎症反应，本项目拟以髓样分化因子(MyD88), 嘌呤受体P2X配体门控离子通道7受体(P2X7R)和细胞因子信号传导抑制蛋白(SOCS1)作为miR-150作用的候选靶标，通过功能获得和功能缺失实验确定其具体靶点及作用机制；在痛风发展不同阶段检测miR-150、miR-150靶标、miR-150靶标相关信号分子及抗炎/致炎因子水平，证实miR-150参与急性痛风自发缓解及作用机制，并初步阐明miR-150在痛风病程发展中的作用。本课题从miRNAs调节方面揭示痛风的病理生理机制，为临床靶向性治疗提供理论基础。

痛风性关节炎是由于尿酸盐晶体沉积在关节及其周围所致的无菌性性关节炎，具有反复的急性发作和自发性缓解的独特特征，但痛风炎症自限的确切分子机制仍不清楚。我们采用实时荧光定量PCR、生物信息学、细胞转染、蛋白印迹，酶联免疫吸附实验、双荧光素酶报告系统。本研究结果提示，miR-150在痛风患者PBMCs中和MSU诱导的巨噬细胞炎症反应中表达异常。进一步通过生物信息学预测并验证了SOCS1为miR-150靶分子；SOCS1在痛风患者和MSU诱导的炎症反应中表达显著降低， 而STAT3、NF-κB p65 mRNA和 p-STAT3 和p-NF-κB p65蛋白水平在痛风患者和MSU诱导的炎症反应中显著增加，并通过转染实验发现SOCS1 mRNA和蛋白水平能够被miR-150抑制，miR-150可能通过下调SOCS1的表达，进而促进NF-κB信号通路和STAT3的活化，使炎症因子分泌增多，进而加重炎症反应。因此，深入研究miRNA参与痛风炎症调控，有助于阐明痛风的发病机制。