Tian Jing, Zhou Dapeng, Xiang Liangbi, Xie Bing, Wang Baichuan, Li Yang, Liu Xinwei
Department of Orthopaedics, General Hospital of Northern Theater Command, Shenyang, China.
Drug Dev Res. 2022 Nov;83(7):1654-1672. doi: 10.1002/ddr.21985. Epub 2022 Sep 7.
Gouty arthritis is an inflammatory disease induced by monosodium urate (MSU), and is closely related to the activation of inflammasomes. Calycosin plays an anti-inflammatory role in arthritis. This study explored the mechanism of Calycosin in MSU-induced gouty arthritis. MSU-induced gouty arthritis mouse models with or without treatment of Calycosin were established, and physiological and pathological indicators were determined. Similarly, peripheral blood mononuclear cells (PBMCs) and THP-1 macrophages were used in vitro. Lactate dehydrogenase (LDH) was tested. The degree of centrifugal infiltration was detected by immunofluorescence. ELISA and quantitative reverse-transcription polymerase chain reaction were conducted to determine the levels of inflammatory factors. Immunohistochemistry, immunofluorescence, and flow cytometry were utilized to detect the content of caspase-1. Protein expressions of NF-κB-, p62-Keap1 pathway-, and pyroptosis-related factors were examined by western blot. In MSU-induced mouse models, calycosin increased mechanical hyperalgesia but decreased the swelling index of the mouse knee joint in a time-dependent manner. MSU treatment increased inflammatory cells and LysM-eGFP neutrophils recruitment in vivo, and promoted the LDH content in vitro, and meanwhile, calycosin reversed the aforementioned effects of MSU. In addition, calycosin repressed the release of inflammatory factors, promoted p62 level and diminished the levels of AIM2, caspase-1, ASC, IL-1β, Keap1, Cleaved GSDMD, and Cleaved caspase-1 and phosphorylation of p65 and IκBα in MSU-induced mouse or cell models. Furthermore, AIM2 silencing also inhibited MSU-induced inflammation and pyroptosis. Collectively, calycosin may inhibit AIM2 inflammasomes-mediated inflammation and pyroptosis through NF-κB and p62-Keap1 pathways, ultimately playing a protective role in gouty arthritis.
痛风性关节炎是一种由尿酸单钠（MSU）诱导的炎症性疾病，与炎性小体的激活密切相关。毛蕊异黄酮在关节炎中发挥抗炎作用。本研究探讨了毛蕊异黄酮在MSU诱导的痛风性关节炎中的作用机制。建立了给予或不给予毛蕊异黄酮治疗的MSU诱导的痛风性关节炎小鼠模型，并测定了生理和病理指标。同样，在体外使用外周血单个核细胞（PBMCs）和THP-1巨噬细胞。检测乳酸脱氢酶（LDH）。通过免疫荧光检测离心浸润程度。进行酶联免疫吸附测定（ELISA）和定量逆转录聚合酶链反应以测定炎症因子水平。利用免疫组织化学、免疫荧光和流式细胞术检测半胱天冬酶-1（caspase-1）的含量。通过蛋白质免疫印迹法检测核因子κB（NF-κB）、p62- Kelch样环氧氯丙烷相关蛋白1（Keap1）途径和细胞焦亡相关因子的蛋白表达。在MSU诱导的小鼠模型中，毛蕊异黄酮增加了机械性痛觉过敏，但以时间依赖性方式降低了小鼠膝关节的肿胀指数。MSU处理增加了体内炎性细胞和溶菌酶M（LysM）-绿色荧光蛋白（eGFP）中性粒细胞的募集，并促进了体外LDH含量，同时，毛蕊异黄酮逆转了MSU的上述作用。此外，毛蕊异黄酮抑制了炎症因子的释放，提高了p62水平，并降低了MSU诱导的小鼠或细胞模型中无黑色素缺失蛋白2（AIM2）、caspase-1、凋亡相关斑点样蛋白（ASC）、白细胞介素-1β（IL-1β）、Keap1、裂解的gasdermin D（GSDMD）、裂解的caspase-1以及p65和IκBα的磷酸化水平。此外，AIM2沉默也抑制了MSU诱导的炎症和细胞焦亡。总体而言，毛蕊异黄酮可能通过NF-κB和p62-Keap1途径抑制AIM2炎性小体介导的炎症和细胞焦亡，最终在痛风性关节炎中发挥保护作用。
