Laboratory of Pain, Inflammation, Neuropathy, and Cancer, Department of Pathology, Londrina State University, Londrina, PR, Brazil.
Departament of Physiological Sciences, Centre of Biological Sciences, Londrina State University, Londrina, PR, Brazil.
Inflammopharmacology. 2020 Aug;28(4):979-992. doi: 10.1007/s10787-020-00686-7. Epub 2020 Feb 11.
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 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 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.
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。然而，其对特定细胞类型和酵母聚糖诱导的炎症的作用尚不清楚。我们旨在评估 HMC 在酵母聚糖诱导的关节炎模型中的作用。进行了 HMC（10、30 或 100mg/kg）的剂量反应曲线以确定在关节内酵母聚糖刺激后最有效的镇痛剂量。使用卡尺测定膝关节肿胀。通过对膝关节冲洗液中的细胞计数以及组织病理学分析来进行白细胞募集。通过比色测定法（GSH、FRAP、ABTS 和 NBT）和 RT-qPCR（gp91 和 HO-1 mRNA 表达）来测量氧化应激。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产生。通过免疫荧光分析体外 NF-κB 激活。我们观察到 HMC 减轻了机械性过敏和膝关节肿胀、白细胞募集和促炎细胞因子水平。我们还观察到酵母聚糖诱导的氧化应激减少，总抗氧化能力增加，gp91 减少，HO-1 mRNA 表达增加。相应地，总 ROS 产生和巨噬细胞 NFκB 激活减少。使用分子对接分析揭示了 HMC 与 NFκB p65 在 Ser276 处的相互作用。因此，本工作中提出的数据表明 HMC 在酵母聚糖诱导的关节炎模型中具有作为镇痛和抗炎药的用途，可能通过靶向巨噬细胞中的 NFκB 激活。
