Silva Thaís Pereira da, Moura Valéria Mourão de, Souza Maria Carolina Scheffer de, Santos Vanessa Neves Carvalho, Silva Kissinara A M Moreira da, Mendes Mara Giselle Guimarães, Nunez Cecília Verônica, Almeida Patrícia Danielle Oliveira de, Lima Emerson Silva, Mourão Rosa Helena Veras, Dos-Santos Maria Cristina
Programa Multi-Institucional de Pós-graduação em Biotecnologia, Laboratório de Imunoquímica, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas, Manaus, AM, Brazil; Laboratório de Imunoquímica, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas, AM, Brazil.
Programa Multi-Institucional de Pós-graduação em Biotecnologia, Laboratório de Imunoquímica, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas, Manaus, AM, Brazil; Laboratório de Imunoquímica, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas, AM, Brazil.
J Ethnopharmacol. 2016 May 13;183:166-175. doi: 10.1016/j.jep.2016.02.043. Epub 2016 Mar 3.
The plant species Connarus favosus is used in folk medicine in the west of Pará state, Brazil, to treat snakebites.
To investigate the potential of the aqueous extract of Connarus favosus (AECf) to inhibit hemorrhagic and phospholipase A2 activities induced by Bothrops atrox venom (BaV) and to determine the antioxidant and antimicrobial potentials of the extract.
AECf was analyzed phytochemically for phenolics (condensed tannins and hydrolyzable tannins) by colorimetry. Antioxidant activity was evaluated by quantitative assays using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Fe(3+)/phenanthroline. Antimicrobial activity was evaluated by the minimal inhibitory concentration test, and cytotoxicity was evaluated using human fibroblast cells (MRC-5). Inhibition of BaV-induced hemorrhagic activity was assessed after oral administration of the extract using pre-treatment, post-treatment and combined (BA plus AECf) treatment protocols. Inhibition of indirect hemolysis caused by phospholipase A2 (PLA2) was investigated in vitro. Interaction between AECf and BaV was investigated by SDS-PAGE electrophoresis, Western blot (Wb) and zymography.
The phytochemical profile of AECf revealed ten secondary metabolite classes, and colorimetry showed high total phenolic and total (condensed and hydrolyzable) tannin content. AECf exhibited high antioxidant and antimicrobial potentials. The IC50 for the cytotoxic effect was 51.91 (46.86-57.50)µg/mL. Inhibition of BaV-induced hemorrhagic activity was significant in all the protocols, and inhibition of PLA2 activity was significant with the two highest concentrations. The BaV/AECf mixture produced the same bands as BaV by itself in SDS-PAGE and Wb although the bands were much fainter. Zymography confirmed the proteolytic activity of BaV, but when the venom was pre-incubated with AECf this activity was blocked.
AECf was effective in reducing BaV-induced hemorrhagic activity when administered by the same route as that used in folk medicine and exhibited antioxidant and antimicrobial potentials.
巴西帕拉州西部的民间医学中使用 Favosus 叶下珠（Connarus favosus）这一植物物种来治疗蛇咬伤。
研究 Favosus 叶下珠水提取物（AECf）抑制具窍蝮蛇毒（BaV）诱导的出血和磷脂酶 A2 活性的潜力，并确定该提取物的抗氧化和抗菌潜力。
采用比色法对 AECf 的酚类物质（缩合单宁和可水解单宁）进行植物化学分析。通过使用 2,2-二苯基-1-苦基肼（DPPH）和铁（Ⅲ）/邻菲啰啉的定量测定来评估抗氧化活性。通过最低抑菌浓度试验评估抗菌活性，并使用人成纤维细胞（MRC-5）评估细胞毒性。在口服提取物后，采用预处理、后处理和联合（BA 加 AECf）处理方案评估对 BaV 诱导的出血活性的抑制作用。在体外研究对磷脂酶 A2（PLA2）引起的间接溶血的抑制作用。通过 SDS-PAGE 电泳、蛋白质印迹（Wb）和酶谱分析研究 AECf 与 BaV 之间的相互作用。
AECf 的植物化学图谱显示有十种次生代谢物类别，比色法显示总酚和总（缩合和可水解）单宁含量较高。AECf 具有较高的抗氧化和抗菌潜力。细胞毒性的 IC50 为 51.91（46.86 - 57.50）μg/mL。在所有方案中，对 BaV 诱导的出血活性的抑制作用均显著，并且在两个最高浓度下对 PLA2 活性的抑制作用显著。在 SDS-PAGE 和 Wb 中，BaV/AECf 混合物产生的条带与 BaV 本身相同，尽管条带要淡得多。酶谱分析证实了 BaV 的蛋白水解活性，但当毒液与 AECf 预孵育时，该活性被阻断。
当按照民间医学中使用的相同途径给药时，AECf 可有效降低 BaV 诱导的出血活性，并具有抗氧化和抗菌潜力。
