Carvalho Raquel, Bonfá Iluska Senna, de Araújo Isaías Muller Jéssica, Pando Silvana Cristina, Toffoli-Kadri Mônica Cristina
Graduate Program in Biotechnology, Faculty of Pharmaceutical Sciences, Food and Nutrition, Federal University of Mato Grosso do Sul (UFMS), Campo Grande, MS, Brazil; Laboratory of Pharmacology and Inflammation, Faculty of Pharmaceutical Sciences, Food and Nutrition, UFMS, Campo Grande, MS, Brazil.
Laboratory of Pharmacology and Inflammation, Faculty of Pharmaceutical Sciences, Food and Nutrition, UFMS, Campo Grande, MS, Brazil.
J Ethnopharmacol. 2023 Jan 10;300:115694. doi: 10.1016/j.jep.2022.115694. Epub 2022 Sep 10.
Libidibia ferrea (Mart. ex. Tul.) L.P. Queiroz is a Brazilian native tree locally known as jucá and pau-ferro, and it has been used in folk medicine for relieving, asthma, bronchitis, sore throat, rheumatism, enterocolitis and fever. The anti-inflammatory properties of L. ferrea were confirmed for its stem, fruit, leaves, bark and seeds extracts, however little is known about the natural compounds that may be associated with that response.
In a normal physiological condition, many enzymes play an important role in catalyzing biological functions. Among them, proteases are of great interest. Although they take part of many biological systems, as the inflammatory process, when deregulated, proteases may cause system malfunctions, such as under- or overproduction of cytokines, or immune cells activation. Thus, protease inhibitors prevent these immune responses by regulating proteases. The objective of this study was to evaluate the anti-inflammatory and anti-nociceptive response of a protease inhibitor purified from L. ferrea seeds (LfTI).
In vitro (5, 50 and 250 μg/mL of LfTI) and in vivo (0.6, 3 e 15 mg/kg of LfTI) assays were performed. Male Swiss mice weighing 18-25 g were used for cell harvesting and for the in vivo assays. The anti-inflammatory activity was analyzed in vitro by macrophage cytotoxicity, hydrogen peroxide (HO) production, and cell adhesion assays; and in vivo by leukocyte recruitment, nitric oxide (NO) production, vascular permeability, paw edema and mast cell degranulation assays. The anti-nociceptive activity was evaluated through abdominal writhing test induced by acetic acid and formalin sensitization.
Our results showed that, in vitro, LfTI is not cytotoxic. Also, LfTI (50 μg/mL) inhibited macrophage HO production (48.2%), and adhesion (48.4%). LfTI (0.6, 3 e 15 mg/kg) decreased polymorphonuclear cell recruitment dose-dependently, and it inhibited NO production (53%), vascular permeability (40.7%) and paw edema at 3 mg/kg at different time, but it did not inhibit mast cell degranulation. Besides, LfTI did not inhibit either the number of writhing or the licking time in the formalin test in the second phase (inflammatory). However, LfTI (3 mg/kg) inhibited licking time at the first phase (neurogenic) in the formalin sensitization (46.1%).
Our results show that LfTI has anti-inflammatory and antinociceptive (neurogenic pain) effects, and these effects might be associated with the inhibition of inflammatory proteases and/or protease-activated receptors activation hindering.
铁刀木（Libidibia ferrea (Mart. ex. Tul.) L.P. Queiroz）是一种原产于巴西的树木，当地称为jucá和pau-ferro，在民间医学中被用于缓解哮喘、支气管炎、喉咙痛、风湿病、小肠结肠炎和发烧。铁刀木的抗炎特性已在其茎、果实、叶子、树皮和种子提取物中得到证实，然而，对于可能与该反应相关的天然化合物知之甚少。
在正常生理条件下，许多酶在催化生物功能中起重要作用。其中，蛋白酶备受关注。尽管它们参与许多生物系统，如炎症过程，但当失调时，蛋白酶可能导致系统功能紊乱，如细胞因子产生不足或过量，或免疫细胞激活。因此，蛋白酶抑制剂通过调节蛋白酶来预防这些免疫反应。本研究的目的是评估从铁刀木种子中纯化的一种蛋白酶抑制剂（LfTI）的抗炎和抗伤害感受反应。
进行了体外（LfTI浓度为5、50和250μg/mL）和体内（LfTI剂量为0.6、3和15mg/kg）试验。体重18 - 25g的雄性瑞士小鼠用于细胞采集和体内试验。通过巨噬细胞细胞毒性、过氧化氢（HO）产生和细胞黏附试验在体外分析抗炎活性；通过白细胞募集、一氧化氮（NO）产生、血管通透性、爪肿胀和肥大细胞脱颗粒试验在体内分析抗炎活性。通过醋酸诱导的腹部扭体试验和福尔马林致敏评估抗伤害感受活性。
我们的结果表明，在体外，LfTI没有细胞毒性。此外，LfTI（50μg/mL）抑制巨噬细胞HO产生（48.2%）和黏附（48.4%）。LfTI（0.6、3和15mg/kg）剂量依赖性地减少多形核细胞募集，并在不同时间抑制3mg/kg剂量下的NO产生（53%）、血管通透性（40.7%）和爪肿胀，但不抑制肥大细胞脱颗粒。此外，LfTI在福尔马林试验的第二阶段（炎症阶段）既不抑制扭体次数也不抑制舔舐时间。然而，LfTI（3mg/kg）在福尔马林致敏的第一阶段（神经源性疼痛阶段）抑制舔舐时间（46.1%）。
我们的结果表明，LfTI具有抗炎和抗伤害感受（神经源性疼痛）作用，这些作用可能与抑制炎性蛋白酶和/或阻碍蛋白酶激活受体的激活有关。
