Asphodelus tenuifolius is traditionally used in the management of rheumatic pain and inflamed body parts. The current study validated its traditional use as an anti-arthritic and anti-inflammatory agent using a series of in vivo models. Carrageenan and histamine-induced acute oedema models were employed to study the effects of n-hexane (n-HeAT) and ethanolic (EeAT) extracts on acute inflammatory mediators and were found to inhibit oedema formation in a dose-dependent manner. Formalin and complete Freund’s adjuvant (CFA) were injected into the hind paw of rats for the induction of arthritis. In the formalin model both n-HeAT and EeAT showed significantly better (p < 0.05) anti-oedema effects from day 6 onward. In CFA model rats were treated on 8th day of induction with extracts at the doses of 250, 500, and 750 mg/kg respectively. Piroxicam (10 mg/kg) and normal saline (10 mL/kg) were used as positive and negative controls respectively. Both n-HeAT and EeAT significantly (p < 0.05) decreased arthritis development in a time-dependent manner and at 28th day extent of inflammation was even less than that observed at day 8. The arthritic score was measured at day 12, 16, 20, 24, and 28 and was observed to be significantly less (p < 0.05) in animals treated with 750 mg/kg of n-HeAT and EeAT, respectively. Joint inflammation (p < 0.01), bone erosion (p < 0.001) and, pannus formation (p < 0.01) were significantly declined in A. tenuifolius treatment groups. Radiographic evaluations (X-ray) were conducted to check bone integrity and extent of inflammation and were observed to be diminished at day 28 in A. tenuifolius extracts treated groups. HPLC was performed to screen the phytochemical profile of n-HeAT and EeAT and were found to contain flavonoids and phenolic compounds. Quantitative real‐time polymerase chain reaction (qPCR) was performed to detect effects of n-HeAT and EeAT treatments on inflammatory markers i.e., IL-4, IL-6, IL-10, COX-2, NF-κB, and I-κB using blood samples. ELISA assays were performed for the detection of levels of C-reactive proteins, respectively. Significant downregulation of TNF-α, IL-4, IL-6, IL-1β, COX-2, NF-κB with simultaneous upregulation of IL-10 and I-κB was observed in n-HeAT and EeAT treatment groups. ELISA assays also showed significant (p < 0.05) down-modulation in the serum levels of CRP and TNF-α. Both extracts showed relatively weak antioxidant activities as compared with ascorbic acid in in vitro assay. Based on findings of the current study it is concluded that A. tenuifolius has anti-inflammatory and anti-arthritic effects and thus has potential to be used as an adjunct to standard NSAIDs therapy.

Graphic abstract

Asphodelus tenuifolius传统上用于治疗风湿性疼痛和发炎的身体部位。目前的研究使用一系列体内模型验证了其作为抗关节炎和抗炎剂的传统用途。角叉菜胶和组胺引起的急性水肿模型来研究的效果Ñ正己烷（正热）和乙醇（EEAT对急性炎症介质）的萃取液，并发现抑制水肿形成以剂量依赖的方式。将福尔马林和完全弗氏佐剂(CFA)注射到大鼠后爪中以诱导关节炎。在福尔马林模型中，n-HeAT和EeAT 都表现出明显更好的效果（p < 0.05) 从第 6 天开始的抗水肿效果。在 CFA 模型中，大鼠在诱导的第 8 天分别以 250、500 和 750 mg/kg 的剂量接受提取物治疗。吡罗昔康 (10 mg/kg) 和生理盐水 (10 mL/kg) 分别用作阳性和阴性对照。两个正热和EEAT显著（p  <0.05）以时间依赖的方式和在炎症第28天下降程度发展关节炎甚至小于在第8天观察到的关节炎得分在第12天，16个，20测量分别 用 750 mg/kg 的n-HeAT和EeAT处理的动物中观察到显着降低（p < 0.05）、24 和 28 。关节炎症（p < 0.01)、骨侵蚀 ( p  < 0.001) 和血管翳形成 ( p  < 0.01) 在A. tenuifolius治疗组中显着下降。进行放射照相评估 (X 射线) 以检查骨完整性和炎症程度，并观察到在第 28 天在细叶松子虫提取物治疗组中减少。进行 HPLC 以筛选n-HeAT和EeAT的植物化学特征，发现它们含有类黄酮和酚类化合物。进行定量实时聚合酶链反应 (qPCR) 以检测n-HeAT和EeAT 的影响使用血液样本对炎症标志物（即 IL-4、IL-6、IL-10、COX-2、NF-κB 和 I-κB）进行治疗。分别进行ELISA测定以检测C反应蛋白的水平。在n-HeAT和EeAT治疗组中观察到 TNF-α、IL-4、IL-6、IL-1β、COX-2、NF-κB 显着下调，同时 IL-10 和 I-κB 上调。ELISA 测定还显示 CRP 和 TNF-α 的血清水平显着 ( p < 0.05) 下调。在体外试验中，与抗坏血酸相比，两种提取物都显示出相对较弱的抗氧化活性。根据目前的研究结果得出的结论是A. tenuifolius 具有抗炎和抗关节炎作用，因此有可能用作标准 NSAIDs 治疗的辅助手段。

图形摘要