Said Khalil, Hamayun Muhammad, Rauf Mamoona, Khan Sumera Afzal, Arif Muhammad, Alrefaei Abdulwahed Fahad, Almutairi Mikhlid H, Ali Sajid
Department of Botany, Abdul Wali Khan University Mardan, Garden Campus, Khyber Pakhtunkhwa, Mardan 23200, Pakistan.
Center of Biotechnology and Microbiology, University of Peshawar, Peshawar 25120, Pakistan.
Curr Pharm Des. 2023;29(41):3324-3339. doi: 10.2174/0113816128290636231129074039.
In the present study, we aimed to investigate the extraction and identification of the potential phytochemicals from the Methanolic Extract of (MEDR) using GC-MS profiling for validating the traditional uses of MEDR its efficacy in inflammations by using and approaches in anti-inflammatory models.
GC-MS analysis confirmed the presence of a total of 59 phytochemical compounds. The human red blood cells (HRBC) membrane stabilization assay and heat-induced hemolysis method were used as anti-inflammatory activity of the extract. The analysis was carried out through the Xylene-induced mice ear oedema method. It was found that MEDR at a concentration of 20 μg, 30 μg, and 40 μg showed 35.45%, 36.01%, and 36.33% protection to HRBC in a hypotonic solution, respectively. At the same time, standard Diclofenac at 30 μg showed 45.31% protection of HRBC in a hypotonic solution.
The extract showed inhibition of 25.32%, 26.53%, and 33.31% cell membrane lysis at heating at 20 μg, 30 μg, and 40 μg, respectively. In comparison, standard Diclofenac at 30 μg showed 50.49% inhibition of denaturation to heat. Methanolic extract of the plant exhibited momentous inhibition in xylene-induced ear oedema in mice treated with 30 μg extract were 47.2%, 63.4%, and 78.8%, while inhibition in mice ear oedema treated with 60 μg extract was 34.7%, 43.05%, 63.21% and reduction in ear thickness of standard drug were 57.3%, 59.54%, 60.42% recorded at the duration of 1, 4 and 24 hours of inflammation. Molecular docking and simulations were performed to validate the anti-inflammatory role of the phytochemicals that revealed five potential phytochemicals Stigmasterol,22,23dihydro, Heptadecane,8methyl, Pimaricacid, Germacrene and 1,3Cyclohexadiene,_5(1,5dimethyl4hexenyl)-2methyl which revealed potential or significant inhibitory effects on cyclooxygenase-2 (COX-2), tumour necrosis factor (TNF-α), and interleukin (IL-6) in the docking analysis.
The outcome of the study signifies that MEDR can offer a new prospect in the discovery of a harmonizing and alternative therapy for inflammatory disease conditions.
在本研究中，我们旨在通过气相色谱 - 质谱联用（GC - MS）分析，从[植物名称]甲醇提取物（MEDR）中提取并鉴定潜在的植物化学物质，以验证MEDR的传统用途，并通过抗炎模型中的[具体方法1]和[具体方法2]方法研究其在炎症中的功效。
GC - MS分析证实共存在59种植物化学化合物。采用人红细胞（HRBC）膜稳定试验和热诱导溶血法评估提取物的抗炎活性。通过二甲苯诱导的小鼠耳肿胀法进行[具体分析方法]分析。结果发现，浓度为20μg、30μg和40μg的MEDR在低渗溶液中对HRBC的保护率分别为35.45%、36.01%和36.33%。同时，30μg的标准双氯芬酸在低渗溶液中对HRBC的保护率为45.31%。
提取物在20μg、30μg和40μg加热条件下，细胞膜裂解抑制率分别为25.32%、26.53%和33.31%。相比之下，30μg的标准双氯芬酸对热变性的抑制率为50.49%。植物甲醇提取物对二甲苯诱导的小鼠耳肿胀有显著抑制作用，30μg提取物处理的小鼠耳肿胀抑制率分别为47.2%、63.4%和78.8%，60μg提取物处理的小鼠耳肿胀抑制率分别为34.7%、43.05%、63.21%，标准药物在炎症1小时、4小时和24小时时耳厚度的减少率分别为57.3%、59.54%、60.42%。进行分子对接和模拟以验证植物化学物质的抗炎作用，结果显示五种潜在的植物化学物质，即豆甾醇、22,23 - 二氢、8 - 甲基十七烷、海松酸、吉马烯和1,3 - 环己二烯，5 - (1,5 - 二甲基 - 4 - 己烯基) - 2 - 甲基，在对接分析中对环氧合酶 - 2（COX - 2）、肿瘤坏死因子（TNF - α）和白细胞介素（IL - 6）具有潜在或显著的抑制作用。
该研究结果表明，MEDR可为发现用于炎症性疾病的协调和替代疗法提供新的前景。
