Wang Yu
Department of Rheumatism Immunology, First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, 150001, People's Republic of China.
Inflamm Res. 2021 Mar;70(3):323-341. doi: 10.1007/s00011-021-01439-0. Epub 2021 Feb 9.
Tripterine (Trip) is frequently applied to alleviate inflammation in various diseases such as rheumatoid arthritis. Macrophages have both anti-inflammatory and pro-inflammatory functions. However, whether Trip can inhibit cell inflammation in gouty arthritis (GA) remains undiscovered and whether the mechanism involved in macrophage polarization is also undetermined. This paper aims to study the effects of Trip on inflammation and macrophage polarization in GA.
Monosodium urate (MSU) crystals were used to establish GA mouse models, and bone marrow-derived macrophages (BMDMs) were induced to construct GA cell models. Pretreatments of Trip and injection of Antagomir-449a/Agomir-449a were performed on mice for 6 days. The effects of Trip and miR-449 on toe swelling, joint damage of GA mouse were examined. The alternations on cell morphology, cell proliferation marker Ki67, inflammatory cytokines, NLRP3 inflammasome, and NF-κB signaling-related proteins were also determined both in vivo and in vitro. Dual-luciferase reporter gene assay and RIP assay were adopted to estimate the targeting relationship between miR-449a and NLRP3.
GA mouse model had increased M1 macrophage, intensified inflammation response, along with suppressed miR-449a expression. Following administration of Trip attenuated cell inflammation, promoted macrophage polarize to M2 phenotype, elevated miR-449a expression, repressed the phosphorylation levels of NF-κB signaling-related proteins, and diminished IκBα expression in vivo and in vitro. However, inhibition of miR-449a hindered the favorable effect of Trip on GA and increased NLRP3 inflammasome expression. MiR-449a directly targeted NLRP3. Overexpression of NLRP3 partially eliminated the biological effects of miR-449a agonist.
Trip regulates macrophage polarization through miR-449a/NLRP3 axis and the STAT3/NF-κB pathway to mitigate GA. The elucidation on the molecular mechanism of Trip in GA may provide theoretical guidance for clinical therapy of GA.
雷公藤红素（Trip）常用于缓解类风湿关节炎等多种疾病的炎症。巨噬细胞具有抗炎和促炎功能。然而，Trip是否能抑制痛风性关节炎（GA）中的细胞炎症尚未可知，其涉及巨噬细胞极化的机制也未明确。本文旨在研究Trip对GA炎症和巨噬细胞极化的影响。
用尿酸钠（MSU）晶体建立GA小鼠模型，诱导骨髓来源的巨噬细胞（BMDM）构建GA细胞模型。对小鼠进行Trip预处理并注射Antagomir - 449a/Agomir - 449a，持续6天。检测Trip和miR - 449对GA小鼠足趾肿胀、关节损伤的影响。还在体内和体外测定细胞形态、细胞增殖标志物Ki67、炎性细胞因子、NLRP3炎性小体和NF - κB信号相关蛋白的变化。采用双荧光素酶报告基因测定法和RIP测定法评估miR - 449a与NLRP3之间的靶向关系。
GA小鼠模型中M1巨噬细胞增加，炎症反应增强，同时miR - 449a表达受到抑制。给予Trip后可减轻细胞炎症，促进巨噬细胞向M2表型极化，提高miR - 449a表达，抑制NF - κB信号相关蛋白的磷酸化水平，并在体内和体外降低IκBα表达。然而，抑制miR - 449a会阻碍Trip对GA的有益作用并增加NLRP3炎性小体表达。miR - 449a直接靶向NLRP3。NLRP3的过表达部分消除了miR - 449a激动剂的生物学效应。
Trip通过miR - 449a/NLRP3轴和STAT3/NF - κB途径调节巨噬细胞极化以减轻GA。对Trip在GA中分子机制的阐明可为GA的临床治疗提供理论指导。
