Abstract：
UNASSIGNED: 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.
OBJECTIVE: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.
OBJECTIVE: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.
摘要：
未经批准：紫藤是西高止山脉的常绿植物，印度,使用传统的部落治疗师，由于其治疗炎症和疼痛缓解程序的优良生物潜力。还消耗树皮提取物以缓解骨骨折部位的炎症变化。印度的传统药用植物必须以其多样化的植物化学成分为特征，它的交互式多个目标站点，揭示生物潜能背后隐藏的分子机制。
目的：该研究集中在植物材料表征上，计算分析(预测研究)，毒理学筛查（体内），以及在LPS诱导的RAW264.7细胞中的红牛甲醇树皮提取物(PRME)的抗炎评价。
方法：使用PRME的纯化合物分离及其生物相互作用来预测生物活性成分，分子靶标,和PRME抑制炎症介质的分子途径。在脂多糖（LPS）诱导的RAW264.7巨噬细胞模型中评估了PRME提取物的抗炎作用。对30只健康的Sprague-Dawley实验大鼠进行了PRME的毒性评估，随机分为五组进行90天的毒理学评价。使用ELISA方法测量组织的氧化应激水平和器官毒性标志物。进行核磁共振波谱(NMR)以表征生物活性分子。
结果：结构表征显示存在香草酸，4-O-甲基没食子酸，E-白藜芦醇,gallocatechin,4'-O-甲基gallocechin，和儿茶素。NF-kB的分子对接表现出与香草酸和4-O-甲基没食子酸的显着相互作用，结合能为-351.159Kcal/Mol和-326.5505Kcal/Mol，分别。PRME处理的动物显示总GPx和抗氧化剂水平(SOD和过氧化氢酶)增加。组织病理学检查显示肝脏没有变化，肾和脾组织的细胞模式。PRME抑制促炎参数(IL-1β，IL-6和TNF-α）在LPS诱导的RAW264.7细胞中。TNF-α和NF-kB蛋白表达研究的蛋白水平显着降低，并与基因表达研究表现出良好的相关性。
结论：本研究确立了PRME作为针对LPS激活的RAW264.7细胞诱导的炎症介质的有效抑制剂的治疗潜力。对SD大鼠的长期毒性评估证实了PRME高达250mg/体重3个月的无毒性质。
目的：该研究集中在植物材料表征上，
方法：使用PRME的纯化合物分离及其生物相互作用来预测生物活性成分，
结果：结构表征显示存在香草酸，
结论：本研究确立了PRME作为针对LPS激活的RAW264.7细胞诱导的炎症介质的有效抑制剂的治疗潜力。
