Sousa Paloma L, Quinet Yves P, Cavalcante Brizeno Luiz André, Sampaio Tiago Lima, Torres Alba Fabíola C, Martins Alice Maria C, Assreuy Ana Maria S
Instituto Superior de Ciências Biomédicas, Universidade Estadual do Ceará (UECE), Av. Paranjana 1700, Itaperi 60 740-000, Fortaleza-CE, Brazil.
Departamento de Análises Clínicas e Toxicológicas, Faculdade de Farmácia, Odontologia e Enfermagem, Universidade Federal do Ceará (UFC), Rua Capitão Francisco Pedro 1210, 60430-370, Fortaleza-CE, Brazil.
Toxicon. 2016 Jul;117:22-9. doi: 10.1016/j.toxicon.2016.03.009. Epub 2016 Mar 23.
Dinoponera quadriceps (Hymenoptera, Formicidae, Ponerinae) is a primitive and endemic ant of Northeastern Brazil, that uses its sting and associated venom gland to capture preys and for defense. Venom of Dinoponera is of potential clinical importance, since it causes intense local pain, accompanied by erythema and edema, when injected by the sting. With other hymenopteran venoms, inflammatory effects are also reported. The aim of this study was to evaluate the inflammatory activity of D. quadriceps venom (DqV) in mice. Acrylamide electrophoresis of DqV revealed five main protein bands varying between 15 and 100 kDa, confirming the proteinous nature of DqV. DqV subplantar injection elicited edema at 5 μg/kg (3 fold), 50 μg/kg (4 fold) or 500 μg/kg (7 fold) from zero to 360 min compared to saline. DqV (50 μg/kg) increased vascular permeability (4 fold) in the first hour after induction. The paw tissue histology showed moderate inflammatory focus caused by DqV (50 μg/kg) in the first hour of paw edema, but severe tissue changes (edema, inflammatory infiltrate and focal areas of hemorrhage) in the third hour. Intraperitoneal injection of DqV (50 μg/kg) stimulated neutrophil (7 fold) and mononuclear (1.4 fold) migration vs saline. DqV edematogenic effect was inhibited by dexamethasone (92%), thalidomide (82%), cyproheptadine (62%), AA861 (58%), celecoxib (34%) or l-NAME (34%), but the neutrophil migration was only by dexamethasone (57%). DqV-elicited neutrophil migration at 50 μg/kg was potentiated 1.7 fold by the animals pre-treatment with 3% thioglycolate. DqV injection increased the levels of interleukin-1 beta (IL-1β) in peritoneal cavities. DqV (50, 100 and 200 μg/mL) increased phospholipase activity (A425nm) from 10 min to 40 min. Raw 267 macrophages incubated with DqV (from 3.12 to 50 mg/mL) showed no significant decrease in cell viability or LDH measurements and at 35 μg/mL induced increase in IL-1β (from 3 to 6 h). This study demonstrated, in mice, the inflammatory effect of D. quadriceps venom, characterized by edema, increase in vascular permeability and neutrophil migration, implying the participation of resident macrophages and IL-1β, among other inflammatory mediators.
四节大齿猛蚁（膜翅目，蚁科，猛蚁亚科）是巴西东北部一种原始的地方性蚂蚁，它利用其螫针和相关的毒腺来捕获猎物和进行防御。大齿猛蚁的毒液具有潜在的临床重要性，因为当通过螫针注入时，它会引起强烈的局部疼痛，并伴有红斑和水肿。与其他膜翅目毒液一样，也有炎症效应的报道。本研究的目的是评估四节大齿猛蚁毒液（DqV）在小鼠中的炎症活性。DqV的丙烯酰胺电泳显示有五条主要蛋白带，分子量在15至100 kDa之间，证实了DqV的蛋白质性质。与生理盐水相比，DqV足底注射5 μg/kg（3倍）、50 μg/kg（4倍）或500 μg/kg（7倍）时，在0至360分钟会引起水肿。DqV（50 μg/kg）在诱导后的第一小时增加了血管通透性（4倍）。爪组织组织学显示，在爪水肿的第一小时，DqV（50 μg/kg）引起中度炎症灶，但在第三小时出现严重的组织变化（水肿、炎症浸润和局部出血区域）。腹腔注射DqV（50 μg/kg）刺激中性粒细胞（7倍）和单核细胞（1.4倍）迁移，而生理盐水组无此现象。DqV的致水肿作用被地塞米松（92%）、沙利度胺（82%）、赛庚啶（62%）、AA861（58%）、塞来昔布（34%）或L - NAME（34%）抑制，但中性粒细胞迁移仅被地塞米松（57%）抑制。用3%硫代乙醇酸盐预处理动物后，50 μg/kg的DqV引起的中性粒细胞迁移增强了1.7倍。DqV注射增加了腹腔中白细胞介素 - 1β（IL - 1β）的水平。DqV（50、100和200 μg/mL）在10分钟至40分钟内增加了磷脂酶活性（A425nm）。用DqV（3.12至50 mg/mL）孵育的Raw 267巨噬细胞在细胞活力或乳酸脱氢酶测量方面没有显著下降，在35 μg/mL时诱导IL - 1β在3至6小时内增加。本研究在小鼠中证明了四节大齿猛蚁毒液的炎症作用，其特征为水肿、血管通透性增加和中性粒细胞迁移，这意味着驻留巨噬细胞和IL - 1β以及其他炎症介质的参与。
