Freitas Raul Sousa, do Val Danielle Rocha, Fernandes Maria Ester Frota, Gomes Francisco Isaac Fernandes, de Lacerda José Thalles Jocelino Gomes, SantiGadelha Tatiane, de Almeida Gadelha Carlos Alberto, de Paulo Teixeira Pinto Vicente, Cristino-Filho Gerardo, Pereira Karuza Maria Alves, de Castro Brito Gerly Anne, Bezerra Mirna Marques, Chaves Hellíada Vasconcelos
Master in Biotechnology, Federal University of Ceará, Avenida Comandante Maurocélio Rocha Pontes, 100 Derby, CEP: 62.042-280 Sobral, Ceará, Brazil.
Northeast Biotechnology Network (Renorbio), Federal University of Pernambuco, Av. Prof. Moraes Rego, 1235 Cidade Universitária, CEP: 50670-901 Recife, Pernambuco, Brazil.
Int Immunopharmacol. 2016 Sep;38:313-23. doi: 10.1016/j.intimp.2016.06.012. Epub 2016 Jun 23.
Temporomandibular joint (TMJ) disorders show inflammatory components, heavily impacting on quality of life. Abelmoschus esculentus is largely cultivated in Northeastern Brazil for medicinal purposes, having it shown anti-inflammatory activity. We evaluated A. esculentus lectin (AEL) efficacy in reducing zymosan-induced temporomandibular joint inflammatory hypernociception in rats along with the mechanism of action through which it exerts anti-inflammatory activity. Animals were pre-treated with AEL (0.01, 0.1 or 1mg/kg) before zymosan (Zy) injection in the TMJ to determine anti-inflammatory activity. To analyse the possible effect of the hemeoxygenase-1 (HO-1) and the nitric oxide (NO) pathways on AEL efficacy, animals were pre-treated with ZnPP-IX (3mg/kg), a specific HO-1 inhibitor, or aminoguanidine (30mg/kg), a selective iNOS inhibitor, before AEL administration. Von Frey test evaluated inflammatory hypernociception, synovial fluid collection was performed to determine leukocyte counting and myeloperoxidase (MPO) activity 6h after Zy injection, and Evans Blue extravasation determined vascular permeability. TMJ tissue was collected for histopathological analysis (H&E) and immunohistochemistry (TNF-α, IL-1β, HO-1). In addition, TMJ tissue and trigeminal ganglion collection was performed for TNF-α and IL-1β dosage (ELISA). AEL increased inflammatory nociceptive threshold, reduced leukocyte influx along with MPO activity, leukocyte influx into the synovial membrane, and Evans Blue extravasation. It promoted HO-1 overexpression whilst decreased TNF-α and IL-1β expression in the TMJ tissue. AEL reduced TNF-α and IL-1β levels in TMJ tissue and trigeminal ganglion. AEL effects, however, were not observed in the presence of ZnPP-IX. These findings suggest that AEL efficacy depends on TNF-α/IL-1β inhibition and HO-1 pathway integrity.
颞下颌关节（TMJ）疾病具有炎症成分，对生活质量有严重影响。秋葵在巴西东北部大量种植用于药用，已显示出抗炎活性。我们评估了秋葵凝集素（AEL）在减轻大鼠体内酵母聚糖诱导的颞下颌关节炎性痛觉过敏方面的功效，以及其发挥抗炎活性的作用机制。在向颞下颌关节注射酵母聚糖（Zy）之前，用AEL（0.01、0.1或1mg/kg）对动物进行预处理，以确定其抗炎活性。为了分析血红素加氧酶-1（HO-1）和一氧化氮（NO）途径对AEL功效的可能影响，在给予AEL之前，先用特异性HO-1抑制剂ZnPP-IX（3mg/kg）或选择性诱导型一氧化氮合酶抑制剂氨基胍（30mg/kg）对动物进行预处理。采用von Frey试验评估炎性痛觉过敏，在注射Zy后6小时收集滑膜液以确定白细胞计数和髓过氧化物酶（MPO）活性，并通过伊文思蓝外渗测定血管通透性。收集颞下颌关节组织进行组织病理学分析（苏木精-伊红染色）和免疫组织化学分析（肿瘤坏死因子-α、白细胞介素-1β、HO-1）。此外，收集颞下颌关节组织和三叉神经节进行肿瘤坏死因子-α和白细胞介素-1β定量分析（酶联免疫吸附测定）。AEL提高了炎性痛觉阈值，减少了白细胞流入以及MPO活性、白细胞向滑膜的流入和伊文思蓝外渗。它促进了HO-1的过度表达，同时降低了颞下颌关节组织中肿瘤坏死因子-α和白细胞介素-1β的表达。AEL降低了颞下颌关节组织和三叉神经节中肿瘤坏死因子-α和白细胞介素-1β的水平。然而，在存在ZnPP-IX的情况下未观察到AEL的作用。这些发现表明，AEL的功效取决于肿瘤坏死因子-α/白细胞介素-1β的抑制和HO-1途径的完整性。
