巨噬细胞焦亡在痛风病理中的作用及清热解毒、利湿化浊法对其的干预机制

Macrophage pyroptosis which mediated by NLRP3 inflammasome is recognized as one of the crucial pathogenesis of gout. In the last six month, many papers which published in Nature, Cell and other top journals，reveal a number of new molecular mechanisms of pyroptosis. pyroptosis "brake" p62, "throttle" NEK7 and "engine" GSDMD and regulation of p62-NEK7-GSDMD axis in the pyroptosis canonical pathway plays an important role. But there isn’t any research report its implications on the pathological process of gout. Our group had launched a randomized controlled trial to verify the method of heat-clearing, detoxifying, dampness-removing and turbidity-dissolving; pyroptosis for the treatment of gout has exact curative effect, can alleviate joint pain, swelling, tenderness and other symptoms, effectively reduce uric acid level in patients’ serum. Animal experiments prove its inhibition to macrophage pyroptosis, and anti-inflammatory and anti-oxidative effects. Therefore, this study using the urate crystal (MSU) induced murine acute gouty arthritis model and MSU-induced macrophage pyroptosis experiments, with CRISPR/cas9 genome editing technology, exploring dead p62-NEK7-GSDMD regulation to pyroptosis by Chinese medicine of heat-clearing, detoxifying, dampness-removing and turbidity-dissolving, in order to find some new targets for the treatment of gout.

巨噬细胞焦亡（pyroptosis）被公认为痛风的重要病理机制之一。近半年来Nature、Cell等期刊多篇论文揭示了焦亡的全新分子机制，堪称重新定义“焦亡”。焦亡“刹车”p62、“油门”NEK7“和“发动机”GSDMD等关键分子构成的p62-NEK7-GSDMD调控轴在焦亡经典通路中发挥重要作用，但尚无研究报告其对痛风病理进程的影响。本项目组以随机对照试验验证了清热解毒、利湿化浊法治疗痛风有确切疗效，可减轻关节疼痛、肿胀、压痛等症状，降低患者血尿酸水平；并以实验证明其抑制巨噬细胞焦亡、抗炎、抗氧化等作用。因此本研究采用尿酸盐晶体诱导的小鼠急性痛风性关节炎模型、尿酸盐晶体诱导的巨噬细胞焦亡模型，利用siRNA沉默NEK7、CRISPR/Cas9基因敲除p62、GSDMD及双敲除等技术，探索清热解毒、利湿化浊法干预痛风的细胞焦亡p62-NEK7-GSDMD调控轴相关机制，揭示痛风治疗新靶点。

巨噬细胞焦亡被公认为痛风的重要病理机制之一。焦亡“刹车”p62、“油门”NEK7“和“发动机”GSDMD等关键分子构成的p62-NEK7-GSDMD调控轴在焦亡经典通路中发挥重要作用，但尚无研究报告其对痛风病理进程的影响。本项目组以随机对照试验验证了清热解毒、利湿化浊法治疗痛风有确切疗效，可减轻关节疼痛、肿胀、压痛等症状，降低患者血尿酸水平；并以实验证明其抑制巨噬细胞焦亡、抗炎、抗氧化等作用。因此本研究采用尿酸盐晶体诱导的小鼠急性痛风性关节炎模型、尿酸盐晶体诱导的巨噬细胞焦亡模型，利用CRISPR/Cas9 基因敲除等技术，实验研究阐示了期p62- NEK7- GSDMD轴及其焦亡通路对痛风的发生、发展具有调控作用，同时验证了清热解毒、利湿化浊立法的秦皮散加味复方可通过调控p62-NEK7-GSDMD轴进而干预痛风性关节炎的病理进展。进而提示了全新的痛风治疗靶点，为进一步深入开发、应用打基础。