Department of Pharmacognosy, School of Pharmaceutical Sciences, Siksha 'O' Anusandhan Deemed to be University, Bhubaneswar, 751003, Odisha, India.
Department of Pharmaceutics, School of Pharmaceutical Sciences, Siksha 'O' Anusandhan Deemed to be University, Bhubaneswar, 751003, Odisha, India.
J Ethnopharmacol. 2024 Jun 28;328:117991. doi: 10.1016/j.jep.2024.117991. Epub 2024 Mar 7.
Glinus oppositifolius (L.) Aug. DC. belongs to the family Molluginaceae, an annual prostrate herb traditionally used to treat inflammations, arthritis, malarial, wounds, fevers, diarrhoea, cancer, stomach discomfort, jaundice, and intestinal parasites. However, the anti-arthritic activity of the aerial part has still not been reported.
To investigate the antioxidant and anti-arthritic activity of G. oppositifolius in Complete Freund's Adjuvant (CFA) induced rats.
The dried aerial parts of this plant material were defatted with n-hexane and extracted by methanol using a soxhlet apparatus. The in vitro anti-arthritic activity of methanolic extract of G. oppositifolius (MEGO) was evaluated in protein denaturation, membrane stabilization, and inhibition of proteinase assay at 25, 50, 100, 200, and 400 μg/ml concentrations. Female Wistar rats were immunized sub-dermally into the right hind paw with 0.1 ml of CFA. Rats were administered with MEGO at doses of 200 and 400 mg/kg once daily for fourteen days after arthritis induction. Assessment of arthritis was performed by measuring paw diameter, arthritic index, arthritic score, body weight, organ weight, and hematological and biochemical parameters, followed by the analysis of pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6), interleukin-1-beta (IL-1β), cyclooxygenase-2 (COX-2), interleukin 13 (IL-13) and interleukin 10 (IL-10) and histopathological study. In vivo antioxidant effect was investigated in enzymatic assays. The presence of phytoconstituents was analyzed by Gas Chromatography-Mass Spectrometry (GC-MS) and Liquid Chromatography-Mass Spectrometry (LC-MS), respectively. In silico molecular docking study of the compounds was carried out against COX-2, IL-1β, IL-6, and TNF-α using AutoDock 4.2 and BIOVIA-Discovery Studio Visualizer software.
MEGO's in vitro anti-arthritic activity showed dose-dependent inhibition of protein denaturation, membrane stabilization, and proteinase inhibition, followed by significant in vivo anti-arthritic activity. The rats treated with MEGO showed tremendous potential in managing arthritis-like symptoms by restoring hematological, biochemical, and histological changes in CFA-induced rats. MEGO (200 and 400 mg/kg) showed a significant alleviation in the levels of hyper expressed inflammatory mediators (TNF-α, IL-1β, and IL-6) and oxidative stress (SOD, CAT, GSH, and LPO) in CFA-induced rats. Spergulagenin-A as identified by LC-MS analysis, exhibited the highest binding affinity against COX-2 (-8.6), IL-1β (7.2 kcal/mol), IL-6 (-7.4 kcal/mol), and TNF-α (-6.5 kcal/mol).
Provided with the comprehensive investigation, methanolic extract of G. oppositifolius against arthritic-like condition is a proof of concept that revalidates its ethnic claim. The presence of Spergulagenin-A might be responsible for the anti-arthritic activity.
菱叶丁香（L.）Aug. DC. 属于豆科，一年生匍匐草本植物，传统上用于治疗炎症、关节炎、疟疾、伤口、发热、腹泻、癌症、胃部不适、黄疸和肠道寄生虫。然而，其地上部分的抗关节炎活性尚未得到报道。
研究菱叶丁香（G. oppositifolius）在完全弗氏佐剂（CFA）诱导的大鼠中的抗氧化和抗关节炎活性。
用正己烷脱脂该植物材料的干燥地上部分，然后用索氏提取器用甲醇提取。在 25、50、100、200 和 400μg/ml 浓度下，评估菱叶丁香（MEGO）的甲醇提取物对蛋白质变性、膜稳定和蛋白酶抑制的体外抗关节炎活性。雌性 Wistar 大鼠通过皮下将 0.1ml CFA 免疫到右后爪中。在关节炎诱导后，大鼠每天以 200 和 400mg/kg 的剂量给予 MEGO，持续 14 天。通过测量爪直径、关节炎指数、关节炎评分、体重、器官重量以及血液学和生化参数来评估关节炎，然后分析促炎细胞因子，如肿瘤坏死因子-α（TNF-α）、白细胞介素 6 (IL-6)、白细胞介素 1-β (IL-1β)、环氧化酶-2 (COX-2)、白细胞介素 13 (IL-13) 和白细胞介素 10 (IL-10)，并进行组织病理学研究。体内抗氧化作用通过酶促试验进行研究。利用气相色谱-质谱联用仪（GC-MS）和液相色谱-质谱联用仪（LC-MS）分别分析植物成分的存在。使用 AutoDock 4.2 和 BIOVIA-Discovery Studio Visualizer 软件对化合物进行 COX-2、IL-1β、IL-6 和 TNF-α 的计算机对接研究。
MEGO 的体外抗关节炎活性显示出剂量依赖性抑制蛋白质变性、膜稳定和蛋白酶抑制，随后显示出显著的体内抗关节炎活性。用 MEGO 治疗的大鼠在恢复 CFA 诱导的大鼠的血液学、生化和组织学变化方面表现出极大的潜力，可以管理关节炎样症状。MEGO（200 和 400mg/kg）在 CFA 诱导的大鼠中显著减轻了过度表达的炎症介质（TNF-α、IL-1β 和 IL-6）和氧化应激（SOD、CAT、GSH 和 LPO）的水平。通过 LC-MS 分析鉴定出的螺旋甾烷醇-A，对 COX-2（-8.6）、IL-1β（7.2kcal/mol）、IL-6（-7.4kcal/mol）和 TNF-α（-6.5kcal/mol）表现出最高的结合亲和力。
通过全面的研究，菱叶丁香的甲醇提取物对关节炎样情况的作用提供了一个概念验证，证明了其民族声称的合理性。螺旋甾烷醇-A 的存在可能是其抗关节炎活性的原因。
