Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100193, China; Xinjiang Institute of Chinese Materia Medica and Ethnodrug, Urumqi, 830002, China.
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100193, China.
J Ethnopharmacol. 2021 Nov 15;280:114134. doi: 10.1016/j.jep.2021.114134. Epub 2021 Apr 20.
The roots of Caragana stenophylla have been used as folk medicine due to the functions of activating blood, diuresis, analgesic and tonicity, especially in treating rheumatoid arthritis and hypertension. However, the anti-rheumatoid arthritis mechanisms and bioactive ingredients have not previously been fully investigated.
The aim of this study was to assess the anti-rheumatoid arthritis effects of the roots of Caragana stenophylla ethanol extract (EC), elucidate its mechanism of action and identify its active substances.
Anti-rheumatoid arthritis activity of EC was assessed using type II-collagen induced arthritis in rats. Arthritis severity was evaluated by foot paw volume, arthritis index, joint swelling degree and histopathology. The serum inflammatory cytokines and matrix metalloproteinases (MMPs) were also detected by immunohistochemical analysis. In addition, the protein expression of IκB, p-IκB, iNOS and COX-2 was analyzed by western blot. RAW 264.7 macrophage cells were employed to assess the anti-inflammatory effects of fractions and compounds in vitro. UPLC-Q-TOF-MS was adopted to appraise the ingredients of the active fraction of the roots of C. stenophylla. Furthermore, various chromatographic techniques and spectroscopic methods were used for isolation and structure elucidation of compounds.
The results showed that EC could reduce type II collagen-induced rheumatoid arthritis model arthritic score and histopathology markedly at dose of 240 mg/kg. Besides, EC could suppress the levels of pro-inflammatory cytokines (IL-1β, IL-6, IL-8, IL-17, and TNF-α) and matrix metalloproteinases (MMP-3, MMP-9), and the expression levels of COX-2, p-IκB and iNOS also were declined. While, the levels of IL-10 and IκB were increased. The ethyl acetate fraction exhibited potent inhibitory effects against nitric oxide production in RAW 264.7 macrophage cells. Eleven main components including 1 flavonoid and 10 oligostilbenes from active fraction were isolated by mass directed chromatographic techniques. Their structures were determined on the basis of various spectroscopic methods and by comparison with the published NMR data.
The roots of C. stenophylla attenuated arthritis severity, restored serum cytokine imbalances by regulating NF-κB signaling pathway in type II collagen-induced rheumatoid arthritis model. Oligostilbenes were essential ingredients in ethyl acetate extract of C. stenophylla roots. Stilbenes and flavonoids should be responsible for its anti-rheumatoid arthritis activities.
锦鸡儿的根被用作民间药物，具有活血、利尿、止痛和收敛的作用，尤其在治疗类风湿性关节炎和高血压方面。然而，其抗类风湿关节炎的机制和生物活性成分尚未得到充分研究。
本研究旨在评估锦鸡儿根乙醇提取物（EC）的抗类风湿关节炎作用，阐明其作用机制并鉴定其活性成分。
采用Ⅱ型胶原诱导的大鼠关节炎模型评估 EC 的抗类风湿关节炎活性。通过足爪体积、关节炎指数、关节肿胀程度和组织病理学评估关节炎严重程度。通过免疫组化分析检测血清炎症细胞因子和基质金属蛋白酶（MMPs）。此外，通过 Western blot 分析 IκB、p-IκB、iNOS 和 COX-2 的蛋白表达。采用 RAW 264.7 巨噬细胞评估各馏分和化合物的体外抗炎作用。采用 UPLC-Q-TOF-MS 评价锦鸡儿根活性部位的成分。此外，还采用各种色谱技术和光谱方法对化合物进行分离和结构鉴定。
结果表明，EC 在 240mg/kg 剂量下可显著降低Ⅱ型胶原诱导的类风湿关节炎模型的关节炎评分和组织病理学表现。此外，EC 可抑制促炎细胞因子（IL-1β、IL-6、IL-8、IL-17 和 TNF-α）和基质金属蛋白酶（MMP-3、MMP-9）水平，并降低 COX-2、p-IκB 和 iNOS 的表达水平，同时增加 IL-10 和 IκB 的水平。乙酸乙酯馏分对 RAW 264.7 巨噬细胞中一氧化氮的产生具有很强的抑制作用。通过质量导向色谱技术从活性部位分离得到 11 种主要成分，包括 1 种黄酮和 10 种寡苯乙烯。根据各种光谱方法并通过与发表的 NMR 数据进行比较，确定了它们的结构。
锦鸡儿根通过调节 NF-κB 信号通路减轻Ⅱ型胶原诱导的类风湿关节炎模型的关节炎严重程度，恢复血清细胞因子失衡。寡苯乙烯是锦鸡儿根乙酸乙酯提取物的主要成分。苯乙烯和黄酮类化合物可能是其抗类风湿关节炎活性的物质基础。
