Department of Orthopaedic, The First People's Hospital of Yongkang, Yongkang, Zhejiang, China.
Department of Pharmacy, The First People's Hospital of Yongkang, Yongkang, Zhejiang, China.
Bioengineered. 2022 May;13(5):13632-13642. doi: 10.1080/21655979.2022.2081756.
This study aimed to explore the effects of plumbagin on rheumatoid arthritis (RA) and its mechanism. The RA cell model was simulated following the treatment of interleukin-1β (IL-1β). After the treatment of various concentrations of plumbagin, the impact of plumbagin on the cell viability was examined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The collagen-induced arthritis (CIA) model was established using the solution of bovine type II collagen. Hematoxylin-eosin staining was used to observe the changes of ankle joint tissue, while enzyme-linked immunosorbent assay and western blot were applied to detect the level of inflammatory cytokines. Plumbagin inhibited the viability of human fibroblast-like synoviocytes (HFLS) at the concentration of 1 ~ 3.5 μM. The inhibitory effect of 1 μM plumbagin on cell proliferation was similar to that of methotrexate, the drug used as the positive control. Plumbagin downregulated the levels of inflammatory cytokines and matrix metalloproteinases (MMPs) in IL-1β-treated HFLS, and suppressed the activation of IκB and nuclear factor kappa-B (NF-κB) as well as the entry of p65 into the nucleus. It was also demonstrated in animal experiments that plumbagin inhibited the activation of NF-κB pathway, down-regulated the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and MMPs, and alleviated joint damage in CIA-modeled mice. Collectively speaking, plumbagin might down-regulate the levels of inflammatory cytokines and MMPs through inhibiting the activation of the NF-κB pathway, thereby attenuating RA-induced damage to cells and joints.: CIA: Collagen-induced arthritis; ELISA: Enzyme-linked immuno sorbent assay; HFLS: Human fibroblast-like synoviocytes; IL-6: Interleukin-6; IL-1β: Interleukin-1β; NF-κB: nuclear factor kappa-B; MTT: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; MMPs: Matrix metalloproteinase; OD: Optical density; RA: Rheumatoid arthritis; SDS: Sodium dodecyl sulfate; SD: Standard deviation; TNF-α: Tumor necrosis factor-α; PVDF: Polyvinylidene fluoride.
本研究旨在探讨白花丹醌对类风湿关节炎（RA）的作用及其机制。采用白细胞介素-1β（IL-1β）处理模拟 RA 细胞模型。用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）检测不同浓度白花丹醌对细胞活力的影响。采用牛Ⅱ型胶原溶液建立胶原诱导性关节炎（CIA）模型。苏木精-伊红（H&E）染色观察踝关节组织变化，酶联免疫吸附试验（ELISA）和蛋白质印迹法（western blot）检测炎症细胞因子水平。白花丹醌在 1 至 3.5μM 浓度下抑制人成纤维样滑膜细胞（HFLS）活力。1μM 白花丹醌对细胞增殖的抑制作用与阳性对照药物甲氨蝶呤相似。白花丹醌下调 IL-1β 处理的 HFLS 中炎症细胞因子和基质金属蛋白酶（MMPs）水平，抑制 IκB 和核因子κB（NF-κB）的激活以及 p65 进入细胞核。动物实验也表明，白花丹醌抑制 NF-κB 通路激活，下调肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）和 MMPs 水平，减轻 CIA 模型小鼠关节损伤。综上所述，白花丹醌可能通过抑制 NF-κB 通路的激活下调炎症细胞因子和 MMPs 水平，从而减轻 RA 诱导的细胞和关节损伤。 CIA：胶原诱导性关节炎；ELISA：酶联免疫吸附试验；HFLS：人成纤维样滑膜细胞；IL-6：白细胞介素-6；IL-1β：白细胞介素-1β；NF-κB：核因子 kappa-B；MTT：3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐；MMPs：基质金属蛋白酶；OD：光密度；RA：类风湿关节炎；SDS：十二烷基硫酸钠；SD：标准偏差；TNF-α：肿瘤坏死因子-α；PVDF：聚偏二氟乙烯。
