Anish Rajamohanan Jalaja, Mohanan Biji, Aswathy Thankamani Ravikumar, Nair Aswathy, Radhakrishnan K V, Rauf Arun A
Department of Biochemistry, University of Kerala, Trivandrum, 695581, India.
Chemical Sciences and Technology Division, CSIR-National Institute for Interdisciplinary Science and Technology (CSIR-NIIST), Thiruvananthapuram, 695019, India; Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201002, India.
J Ethnopharmacol. 2023 May 23;308:116262. doi: 10.1016/j.jep.2023.116262. Epub 2023 Feb 14.
Pterospermum rubiginosum is an evergreen plant in Western Ghats, India, used by traditional tribal healers due to its excellent biological potential for treating inflammation and pain relief procedures. The bark extract is also consumed to relieve the inflammatory changes at the bone fractured site. The traditional medicinal plant in India have to be characterized for its diverse phytochemical moieties, its interactive multiple target sites, and to reveal the hidden molecular mechanism behind the biological potency.
The study focussed on plant material characterization, computational analysis (prediction study), toxicological screening (In vivo), and anti-inflammatory evaluation of P. rubiginosum methanolic bark extracts (PRME) in LPS-induced RAW 264.7 cells.
The pure compound isolation of PRME and their biological interactions were used to predict the bioactive components, molecular targets, and molecular pathways of PRME in inhibiting inflammatory mediators. The anti-inflammatory effects of PRME extract were evaluated in the lipopolysaccharide (LPS)-induced RAW264.7 macrophage cell model. The toxicity evaluation of PRME was performed in healthy 30 Sprague-Dawley experimental rats, were randomly divided into five groups for toxicological evaluation for 90 days. The tissue levels of oxidative stress and organ toxicity markers were measured using the ELISA method. Nuclear magnetic resonance spectroscopy (NMR) was performed to characterize the bioactive molecules.
Structural characterization revealed the presence of vanillic acid, 4-O-methyl gallic acid, E-resveratrol, gallocatechin, 4'-O-methyl gallocatechin, and catechin. Molecular docking of NF-kB exhibited significant interactions with vanillic acid and 4-O-methyl gallic acid with binding energy -351.159 Kcal/Mol and -326.5505 Kcal/Mol, respectively. The PRME-treated animals showed an increase in total GPx and antioxidant levels (SOD and catalase). Histopathological examination revealed no variation in the liver, renal and splenic tissue's cellular pattern. PRME inhibited the pro-inflammatory parameters (IL-1β, IL-6, and TNF-α) in LPS-induced RAW 264.7 cells. The protein level of TNF-α and NF-kB protein expression study brought out a notable reduction and exhibited a good correlation with the gene expression study.
The current study establishes the therapeutic potential of PRME as an effective inhibitory agent against LPS-activated RAW 264.7 cells induced inflammatory mediators. Long-term toxicity evaluation on SD rats confirmed the non-toxic nature of PRME up to 250mg/body weight for 3 months.
糙毛翅子树是印度西高止山脉的一种常绿植物，传统部落治疗师因其在治疗炎症和缓解疼痛方面具有出色的生物学潜力而使用它。树皮提取物也被用于缓解骨折部位的炎症变化。印度的传统药用植物必须对其多样的植物化学成分、相互作用的多个靶点进行表征，并揭示其生物活性背后隐藏的分子机制。
本研究聚焦于糙毛翅子树甲醇树皮提取物（PRME）的植物材料表征、计算分析（预测研究）、毒理学筛选（体内）以及在脂多糖（LPS）诱导的RAW 264.7细胞中的抗炎评估。
利用PRME的纯化合物分离及其生物相互作用来预测PRME抑制炎症介质的生物活性成分、分子靶点和分子途径。在脂多糖（LPS）诱导的RAW264.7巨噬细胞模型中评估PRME提取物的抗炎作用。在30只健康的Sprague-Dawley实验大鼠中进行PRME的毒性评估，将其随机分为五组进行90天的毒理学评估。使用酶联免疫吸附测定（ELISA）方法测量氧化应激和器官毒性标志物的组织水平。进行核磁共振光谱（NMR）以表征生物活性分子。
结构表征显示存在香草酸、4-O-甲基没食子酸、白藜芦醇、表儿茶素、4'-O-甲基表儿茶素和儿茶素。NF-κB的分子对接显示与香草酸和4-O-甲基没食子酸有显著相互作用，结合能分别为-351.159千卡/摩尔和-326.5505千卡/摩尔。经PRME处理的动物总谷胱甘肽过氧化物酶（GPx）和抗氧化水平（超氧化物歧化酶（SOD）和过氧化氢酶）有所增加。组织病理学检查显示肝脏、肾脏和脾脏组织的细胞模式无变化。PRME在LPS诱导的RAW 264.7细胞中抑制促炎参数（白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α））。TNF-α的蛋白质水平和NF-κB蛋白质表达研究显示显著降低，并且与基因表达研究具有良好的相关性。
当前研究确立了PRME作为一种有效抑制剂对LPS激活的RAW 264.7细胞诱导的炎症介质的治疗潜力。对SD大鼠的长期毒性评估证实，在3个月内，PRME剂量高达250毫克/体重时无毒。
