Department of Pharmacology, Faculty of Pharmacy, The Islamia University of Bahawalpur, Punjab, Pakistan.
Faculty of Pharmaceutical Sciences, Government College University Faisalabad, Punjab, Pakistan.
Anticancer Agents Med Chem. 2021;21(11):1460-1471. doi: 10.2174/1871520620666201005100132.
Inflammation is a vital reaction of the natural immune system that protects against encroaching agents. However, uncontrolled inflammation can lead to complications. Trigonella foenumgraecum is traditionally used as an anti-inflammatory herb.
The current study was conducted to explore the antioxidant, anti-inflammatory, and antiangiogenic potentials of Trigonella foenum-graecum seeds oil.
Oil was extracted from seeds of Trigonella foenum-graecum by cold press method and labelled as TgSO. Phytochemical (GC-MS, Folin-Ciocalteu method) and metal analyses were conducted to evaluate the metalo-chemical profile of TgSO. In vitro antioxidant assays (2,2-diphenyl-1-picrylhydrazyl, 2,2'-azino-bis-3- ethylbenzothiazoline-6-sulfonic acid and ferric reducing antioxidant power) were performed to assess its antioxidant potential. In vitro antimicrobial activity was evaluated using agar disc diffusion method and the safety profile of TgSO was assessed in acute toxicological studies following OECD 425 guidelines. In vivo antiinflammatory activities of TgSO were assessed in carrageenan, serotonin, histamine, formalin, and cotton pelletinduced oedema models. Serum TNF-α, Superoxide Dismutase (SOD) and, Catalases (CAT) levels were assessed by ELISA kits. In vivo antiangiogenic activity of TgSO was screened in chick Chorioallantoic Membrane (CAM) assay. Histopathological studies using excised paws were conducted to observe the effects of TgSO treatment at the tissue level. In silico docking studies were conducted to screen the binding potentials of identified compounds with TNF-α.
Extraction by cold press method yielded 16% of TgSO. Phytochemical analysis of TgSO through GCMS showed the presence of eugenol, dihydrocoumairn, heptadecanoic acid, tri- and tetradecanoic acid, and hexadecanoic acid, respectively. Total phenolic contents of TgSO were found to be 0.30±0.01mg/g gallic acid equivalent in Folin-Ciocalteu method. Metal analysis indicated the presence of different metals in TgSO. Findings of antioxidant models showed the moderate antioxidant potential of TgSO. Findings of antimicrobial assays showed that TgSO was active against bacterial (S. aureus, S. epidermidis) and fungal (C. albicans, and A. niger) strains. In vivo toxicity study data showed that TgSO was safe up to the dose of 5000 mg/kg. Data of oedema models showed a significant (p<0.05) reduction in oedema development in TgSO treated animals in both acute and chronic models. Histopathological evaluations of paws showed minimum tissue infiltration with inflammatory cells in TgSO-treated animals. Treatment with TgSO also significantly (p<0.05) down-regulated TNF-α in serum while levels of SOD and CAT were up-regulated. Findings of the CAM assay revealed the antiangiogenic activity of TgSO. Findings of in silico docking studies showed that identified phytoconstituents can bind with culprit cytokine (TNF-α).
Data obtained from the current study conclude that TgSO has antioxidant, anti-inflammatory, and antiangiogenic effects that validate its traditional uses. Synergistic actions of different phytoconstituents are proposed to be responsible for the observed effects.
炎症是天然免疫系统的重要反应，可抵御入侵物。然而，失控的炎症可能会导致并发症。葫芦巴是一种传统的抗炎草药。
本研究旨在探索葫芦巴种子油的抗氧化、抗炎和抗血管生成潜力。
采用冷榨法从葫芦巴种子中提取油，并标记为 TgSO。通过 GC-MS（福林-考尔考特法）和金属分析对 TgSO 的金属化学特征进行了评估。采用 2,2-二苯基-1-苦基肼基（2,2'-二氮杂双-3-乙基苯并噻唑-6-磺酸和铁还原抗氧化能力）体外抗氧化测定法评估其抗氧化潜力。采用琼脂圆盘扩散法评估其体外抗菌活性，并按照 OECD 425 指南评估 TgSO 的急性毒性研究中的安全性概况。采用角叉菜胶、血清素、组织胺、甲醛和棉纤维球诱导的水肿模型评估 TgSO 的体内抗炎活性。采用 ELISA 试剂盒测定血清肿瘤坏死因子-α（TNF-α）、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）水平。采用鸡绒毛尿囊膜（CAM）试验筛选 TgSO 的体内抗血管生成活性。通过切除爪子进行组织学研究，观察 TgSO 治疗在组织水平上的效果。通过计算机对接研究筛选鉴定化合物与 TNF-α的结合潜力。
采用冷榨法提取 TgSO 的得率为 16%。通过 GC-MS 对 TgSO 的植物化学成分分析表明，分别存在丁香酚、二氢香豆素、十七烷酸、三和十四烷酸以及十六烷酸。采用福林-考尔考特法测定 TgSO 的总酚含量为 0.30±0.01mg/g 没食子酸当量。金属分析表明 TgSO 中存在不同的金属。抗氧化模型的研究结果表明 TgSO 具有中等的抗氧化潜力。抗菌试验结果表明 TgSO 对细菌（金黄色葡萄球菌、表皮葡萄球菌）和真菌（白色念珠菌、黑曲霉）均有活性。体内毒性研究数据表明 TgSO 安全剂量高达 5000mg/kg。水肿模型的数据表明 TgSO 可显著（p<0.05）减少急性和慢性模型中动物的水肿发展。爪子的组织学评估表明 TgSO 治疗的动物中炎性细胞的组织浸润最小。用 TgSO 治疗还显著（p<0.05）下调了血清中的 TNF-α，同时上调了 SOD 和 CAT 的水平。CAM 试验的研究结果显示 TgSO 具有抗血管生成活性。计算机对接研究的结果表明鉴定出的植物化学成分可以与罪魁祸首细胞因子（TNF-α）结合。
本研究得出的数据表明，TgSO 具有抗氧化、抗炎和抗血管生成作用，验证了其传统用途。提出不同植物成分的协同作用可能是观察到的作用的原因。
