江南卷柏黄酮调控NLRP3炎症小体抑制细胞焦亡治疗痛风的作用机制

Gout has become a common and frequently-occurring disease that seriously endangers human health. Relevant studies have confirmed that the key mechanism of gout is the activation of NLRP3 inflammasome. Selaginella moelledorfii, which is commonly used in ethnic medicine of Tujia in the west of Hubei province, has the effect of clearing away heat and detoxification, removing swelling and dampness, and stopping bleed, and it could be used for a variety of acute and chronic inflammation. Our previous studies have found that Selaginella moelledorfii has anti-gout effect, and flavonoids are the main effective components. The flavonoids could inhibit NLRP3 inflammasome activation and pyroptosis, but the mechanism is unclear. Therefore, on the basis of previous studies, this research intends to study the anti-gout activity of flavonoids via inhibiting cell pyrosis in animal model. At the molecular and cellular level, the effects of total flavonoids and major components are also investigated on the regulation of NLRP3 inflammasome signaling pathway in order to investigate anti-gout mechanism of Selaginella moelledorfii comprehensively.

痛风已成为严重危害人类健康的常见病和多发病，已有研究证实痛风发作的核心机制是NLRP3炎症小体活化。江南卷柏为鄂西土家族常用民族药，具有清热解毒、利湿消肿、止血功效，可用于多种急慢性炎症。前期研究发现江南卷柏具有抗痛风作用，黄酮类化合物为其主要有效成分，可抑制NLRP3炎症小体活化和抑制巨噬细胞焦亡，但其具体机理尚不清楚。故本课题拟在前期研究基础上，运用整体动物模型，从抑制细胞焦亡的角度研究江南卷柏黄酮的抗痛风活性；并从分子和细胞水平上，探讨江南卷柏总黄酮和主要成分对NLRP3炎症小体信号通路调控的作用靶标，综合诠释江南卷柏黄酮抗痛风的作用机制。

痛风已成为严重危害人类健康的常见病和多发病，已有研究证实痛风发作的核心机制是NLRP3炎症小体活化。本项目采用LPS诱导RAW264.7细胞和MSU诱导小鼠急性痛风模型，筛选出江南卷柏总黄酮、芹菜素和穗花杉双黄酮具有明显的抗痛风作用，作用机制与其保护细胞膜完整性，改善细胞形态，降低炎症因子释放水平，调控NLRP3/ASC/Caspase-1轴上相关蛋白和mRNA的表达等有关。已从江南卷柏中分离提取出11个双黄酮化合物，采用LPS+ATP/Nigericin/MSU诱导的NLRP3炎症小体活化模型比较其活性，穗花杉双黄酮可更稳定的显著抑制LDH释放，可通过恢复Ca2+稳态，抑制ROS和NLRP3信号通路等途径抑制NLRP3炎症小体的激活从而抑制细胞焦亡。而江南卷柏黄酮苷类化合物可通过ERa激活MAPK和PI3K/Akt/mTOR信号通路，发挥类雌激素活性。本项目阐明了江南卷柏黄酮抗痛风作用的机制，为开发成新的治疗痛风的药物奠定了基础。