da Silva Girlaine Pereira, Fernandes Daniele Corrêa, Vigliano Mariana Vieira, da Fonseca Eduardo Nunes, Santos Shirley Vânia Moura, Marques Paulo Roberto, Justo Maria da Graça, Sabino Kátia Costa de Carvalho, Coelho Marsen Garcia Pinto
Departamento de Bioquímica, Instituto de Biologia Roberto Alcantara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brazil.
Departamento de Biologia Vegetal, Instituto de Biologia Roberto Alcantara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brazil.
J Pharm Pharmacol. 2016 Dec;68(12):1584-1596. doi: 10.1111/jphp.12620. Epub 2016 Oct 10.
Echinodorus macrophyllus (Kunth) Micheli (Alismataceae) is popularly used as an infusion to treat inflammatory diseases. This work fractionated the aqueous extract of E. macrophyllus (AEEm) to improve its anti-inflammatory effects.
Aqueous extract of E. macrophyllus was fractionated by Sephadex LH-20 and analysed by HPLC-DAD. Anti-inflammatory action was evaluated, in vivo, by air pouch model (total leucocyte, protein and leukotriene B (LTB )), and, in vitro, by neutrophil migration (transwell assay) and its Mac1 expression (flow cytometry), and RAW 264.7 nitric oxide (NO) production (Griess reaction).
Fr20 reduced total leucocyte at 2.5 mg/kg (29.7%) while ethanolic extract of E. macrophyllus (EAEm) increased it (94.0%). Fr20 showed higher (P < 0.05) inhibition (89.8%) of LTB in exudate than EAEm (75.0%). Fr20 and EAEm decreased exudate protein and inflammatory infiltrate in pouch tissues, in-vitro neutrophil migration, and NO production. Otherwise, Fr40 did not reduce leucocytes and exudate protein (until 50 mg/kg) nor tissue inflammation, and increased in-vitro NO production. The inhibition of neutrophil migration by EAEm, but not Fr20, was dependent on reduced Mac-1 expression.
The fractionation of AEEm provided a more potent anti-inflammatory fraction containing flavonoids (Fr20) that reduces the migration of neutrophils and LTB4 release, probably contributing to its mechanism of action.
大叶皇冠草(Echinodorus macrophyllus (Kunth) Micheli,泽泻科)通常被用作浸剂来治疗炎症性疾病。本研究对大叶皇冠草水提取物(AEEm)进行分离,以增强其抗炎作用。
采用葡聚糖凝胶LH - 20对大叶皇冠草水提取物进行分离,并通过高效液相色谱 - 二极管阵列检测器(HPLC - DAD)进行分析。在体内,通过气袋模型(检测总白细胞、蛋白质和白三烯B4(LTB4))评估其抗炎作用;在体外,通过中性粒细胞迁移实验(Transwell实验)及其Mac1表达(流式细胞术),以及RAW 264.7细胞一氧化氮(NO)生成实验(Griess反应)进行评估。
Fr20在2.5 mg/kg剂量时可使总白细胞减少29.7%,而大叶皇冠草乙醇提取物(EAEm)却使其增加94.0%。Fr20对渗出液中LTB4的抑制率(89.8%)高于EAEm(75.0%)(P < 0.05)。Fr20和EAEm均可降低渗出液中的蛋白质含量以及气袋组织中的炎症浸润、体外中性粒细胞迁移和NO生成。否则,Fr40在高达50 mg/kg剂量时既不能减少白细胞和渗出液中的蛋白质含量,也不能减轻组织炎症,反而会增加体外NO生成。EAEm对中性粒细胞迁移的抑制作用依赖于Mac - 1表达的降低,而Fr20则不然。
AEEm的分离得到了一种更有效的抗炎组分(Fr20),该组分含有黄酮类化合物,可减少中性粒细胞的迁移和LTB4的释放,这可能是其作用机制的一部分。