Arthritis can be defined as a painful musculoskeletal disorder that affects the joints. Hesperidin methyl chalcone (HMC) is a flavonoid with analgesic, anti-inflammatory, and antioxidant effects. However, its effects on a specific cell type and in the zymosan-induced inflammation are unknown. We aimed at evaluating the effects of HMC in a zymosan-induced arthritis model. A dose–response curve of HMC (10, 30, or 100 mg/kg) was performed to determine the most effective analgesic dose after intra-articular zymosan stimuli. Knee joint oedema was determined using a calliper. Leukocyte recruitment was performed by cell counting on knee joint wash as well as histopathological analysis. Oxidative stress was measured by colorimetric assays (GSH, FRAP, ABTS and NBT) and RT-qPCR (gp91^phox and HO-1 mRNA expression) performed. In vitro, oxidative stress was assessed by DCFDA assay using RAW 264.7 macrophages. Cytokine production was evaluated in vivo and in vitro by ELISA. In vitro NF-κB activation was analysed by immunofluorescence. We observed HMC reduced mechanical hypersensitivity and knee joint oedema, leukocyte recruitment, and pro-inflammatory cytokine levels. We also observed a reduction in zymosan-induced oxidative stress as per increase in total antioxidant capacity and reduction in gp91^phox and increase in HO-1 mRNA expression. Accordingly, total ROS production and macrophage NFκB activation were diminished. HMC interaction with NFκB p65 at Ser276 was revealed using molecular docking analysis. Thus, data presented in this work suggest the usefulness of HMC as an analgesic and anti-inflammatory in a zymosan-induced arthritis model, possibly by targeting NFκB activation in macrophages.

中文翻译：

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橙皮苷甲基查尔酮与NFκBSer276相互作用并抑制酵母聚糖诱导的关节痛和炎症以及RAW 264.7巨噬细胞的活化。

关节炎可以定义为一种影响关节的疼痛性肌肉骨骼疾病。橙皮苷甲基查尔酮（HMC）是一种具有镇痛，抗炎和抗氧化作用的类黄酮。但是，其对特定细胞类型和酵母聚糖诱导的炎症的影响尚不清楚。我们旨在评估HMC在酵母聚糖诱导的关节炎模型中的作用。进行HMC（10、30或100 mg / kg）的剂量反应曲线，以确定关节内酵母聚糖刺激后最有效的镇痛剂量。使用卡尺确定膝关节水肿。通过在膝关节冲洗液上进行细胞计数以及组织病理学分析来进行白细胞募集。通过比色测定（GSH，FRAP，ABTS和NBT）和RT-qPCR（gp91 ^phox）测量氧化应激和HO-1 mRNA表达）。在体外，使用RAW 264.7巨噬细胞通过DCFDA测定评估氧化应激。通过ELISA在体内和体外评估细胞因子的产生。通过免疫荧光分析了体外NF-κB的活化。我们观察到HMC降低了机械性超敏反应和膝关节水肿，白细胞募集以及促炎性细胞因子水平。我们还观察到随着总抗氧化剂能力的增加和gp91 ^phox的减少，酵母聚糖诱导的氧化应激的减少并增加HO-1 mRNA表达。因此，总的ROS产生和巨噬细胞NFκB活化减少。使用分子对接分析揭示了HMC与Ser276上的NFκBp65的相互作用。因此，这项工作中提供的数据表明，HMC在酵母聚糖诱导的关节炎模型中作为镇痛药和抗炎药的作用可能是通过靶向巨噬细胞中的NFκB活化。