Inflammation and angiogenesis are two major contributors to tumourigenesis. Melilotus indicus is traditionally used as an anti-inflammatory agent. The current study was designed to investigate the anti-inflammatory and anti-angiogenic properties of ethanolic extract of M. indicus (Miet) whole plant and its marker compound (coumarin) using a series of in vivo methods. Extraction by maceration was adopted to prepare ethanolic extract. Phytochemical compounds present in Miet were investigated using both qualitative and quantitative methods. In vivo safety profile of Miet was investigated in behavioural studies. Four acute oedema models such as carrageenan, serotonin, histamine-induced paw oedema and xylene-induced ear oedema, and chronic formaldehyde-induced paw oedema model were employed to explore the anti-inflammatory potential of Miet. Chorioallantoic chick membrane assay (CAM) was performed to explore anti-angiogenic potential of Miet. Histopathological evaluations were conducted to access improvement in skin texture of paws. TNF-α ELISA kit was used to study effects of treatment on serum levels of TNF-α. Extraction by maceration resulted in formation of greenish coloured semisolid extract with a high coumarin content. In vivo toxicological studies revealed LD50 of Miet was greater than 8000 mg/kg. Data of acute inflammatory models depicted significant (p < 0.05) inhibition of oedema in Miet, coumarin and standard (piroxicam/indomethacin) treated groups. 750 mg/kg of Miet induced comparable (p > 0.05) anti-inflammatory effects to that of standard-treated groups. Coumarin showed better anti-inflammatory effects in carrageenan-induced paw oedema model as compared with histamine- and serotonin-induced oedema models. Data of chronic inflammatory models also depicted dose-dependent anti-inflammatory attributes of Miet which were comparable with standard treated groups. Significant (p > 0.05) downregulation of TNF-α in serum samples of animals treated with Miet and piroxicam was observed as compared with control group. Furthermore, Miet significantly halted blood vessels formation in CAM assay. Overall, data of the current study highlight that M. indicus has anti-inflammatory and anti-angiogenic potentials, and, thus, can potentially be used as an adjuvant therapy in solid tumours management.

中文翻译：

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研究富含香豆素的 Melilotus indicus 乙醇提取物的体内抗炎和抗血管生成特性。

炎症和血管生成是肿瘤发生的两个主要因素。Melilotus indicus 传统上用作抗炎剂。本研究旨在使用一系列 体内 方法研究 M. indicus (Miet) 全株乙醇提取物及其标记化合物 (香豆素) 的抗炎和抗血管生成特性。采用浸渍法制备乙醇提取物。使用定性和定量方法研究了 Miet 中存在的植物化学化合物。在行为研究中对 Miet 的体内安全性进行了调查。采用角叉菜胶、5-羟色胺、组胺诱导的足部水肿和二甲苯诱导的耳部水肿等四种急性水肿模型，以及慢性甲醛诱导的足部水肿模型来探索 Miet 的抗炎潜力。进行绒毛膜尿囊膜测定 (CAM) 以探索 Miet 的抗血管生成潜力。进行组织病理学评估以改善爪子的皮肤质地。TNF-α ELISA试剂盒用于研究治疗对血清TNF-α水平的影响。浸渍提取导致形成具有高香豆素含量的绿色半固体提取物。体内毒理学研究显示 Miet 的 LD50 大于 8000 mg/kg。急性炎症模型的数据描述了 Miet、香豆素和标准（吡罗昔康/吲哚美辛）治疗组对水肿的显着抑制（p < 0.05）。750 mg/kg Miet 诱导的抗炎作用与标准治疗组相当（p > 0.05）。与组胺和血清素诱导的水肿模型相比，香豆素在角叉菜胶诱导的爪水肿模型中显示出更好的抗炎作用。慢性炎症模型的数据还描述了 Miet 的剂量依赖性抗炎特性，与标准治疗组相当。与对照组相比，观察到用 Miet 和吡罗昔康治疗的动物血清样品中 TNF-α 的显着下调（p > 0.05）。此外，Miet 在 CAM 测定中显着阻止了血管形成。总体而言，当前研究的数据强调了印度支原体具有抗炎和抗血管生成的潜力，因此有可能用作实体瘤管理的辅助治疗。慢性炎症模型的数据还描述了 Miet 的剂量依赖性抗炎特性，与标准治疗组相当。与对照组相比，观察到用 Miet 和吡罗昔康治疗的动物血清样品中 TNF-α 的显着下调（p > 0.05）。此外，Miet 在 CAM 测定中显着阻止了血管形成。总体而言，当前研究的数据强调了印度支原体具有抗炎和抗血管生成的潜力，因此有可能用作实体瘤管理的辅助治疗。慢性炎症模型的数据还描述了 Miet 的剂量依赖性抗炎特性，与标准治疗组相当。与对照组相比，观察到用 Miet 和吡罗昔康治疗的动物血清样品中 TNF-α 的显着下调（p > 0.05）。此外，Miet 在 CAM 测定中显着阻止了血管形成。总体而言，当前研究的数据强调了印度支原体具有抗炎和抗血管生成的潜力，因此有可能用作实体瘤管理的辅助治疗。Miet 在 CAM 测定中显着阻止了血管形成。总体而言，当前研究的数据强调了印度支原体具有抗炎和抗血管生成的潜力，因此有可能用作实体瘤管理的辅助治疗。Miet 在 CAM 测定中显着阻止了血管形成。总体而言，当前研究的数据强调了印度支原体具有抗炎和抗血管生成的潜力，因此有可能用作实体瘤管理的辅助治疗。