Furtado Allanny A, Torres-Rêgo Manoela, Lima Maíra C J S, Bitencourt Mariana A O, Estrela Andréia Bergamo, Souza da Silva Nayara, da Silva Siqueira Emerson Michell, Tomaz José Carlos, Lopes Norberto Peporine, Silva-Júnior Arnóbio Antônio, Zucolotto Silvana M, Fernandes-Pedrosa Matheus F
Laboratório de Tecnologia e Biotecnologia Farmacêutica, Departamento de Farmácia, Faculdade de Farmácia do Rio Grande do Norte-UFRN, Natal, RN, Brazil.
Laboratório de Farmacognosia, Departamento de Farmácia, Faculdade de Farmácia do Rio Grande do Norte-UFRN, Natal, RN, Brazil.
J Ethnopharmacol. 2016 Nov 4;192:225-235. doi: 10.1016/j.jep.2016.07.048. Epub 2016 Jul 19.
Ipomoea asarifolia (Desr.) Roem. and Schult.(Convolvulaceae), popularly known as salsa or salsa-brava, is a plant of which the decoction of leaves is used in folk medicine to treat various inflammatory disorders such of dermatitis, scabies, symptoms of syphilis, skin ulcers and external wounds. However, little is known about possible compounds and mechanisms of action of the plant to support the activities reported by popular use.
The study aimed to identify bioactive molecules present in the crude extract of I. asarifolia leaves and investigate the anti-inflammatory potential of this extract in different experimental in vivo models to improve the understanding on that activity.
Aqueous extract of I. asarifolia leaves was prepared by decoction (1:10 m/v) and its chromatographic profile was obtained by high performance liquid chromatography coupled with diode array detector (HPLC-DAD) and liquid chromatography diode array detector coupled with mass spectrometry (LC-DAD-MS). The potential anti-inflammatory activity of the extract was assessed using the following in vivo models: xylene-induced ear edema (20, 30 and 40mg/kg), evaluating the degree of edema formation; carrageenan-induced peritonitis (10, 20 and 30mg/kg), evaluating leukocyte migration and cytokine levels (IL-1β, IL-6, IL-12 and TNF-α) at 4h; zymosan-induced air pouch inflammation (20, 30 and 40mg/kg), evaluating the kinetics of leukocyte migration by total and differential counts at 6, 24 and 48h. The same tests were conducted using pure compounds identified in the aqueous extract from I. asarifolia leaves in different doses for each experimental model.
The compounds identified in the aqueous extract of I. asarifolia leaves by HPLC-DAD and LC-DAD-MS were rutin, chlorogenic acid and caffeic acid. The extract significantly reduced ear edema induced by xylene (81%, 85% and 86% for doses of 20, 30 and 40mg/kg, respectively, p<0.001), as well as cell migration in experimental models of peritonitis (70%, 78% and 83% for doses of 10, 20 and 30mg/kg, respectively, p<0.001) and air pouch inflammation (58%, 67% and 53% for doses of 20, 30 and 40mg/kg, respectively, p<0.001). In addition, the extract demonstrated the ability to significantly inhibit the production of cytokines IL-1β, IL-6, IL-12 and TNF-α (p<0.001). The secondary metabolites tested (rutin, chlorogenic acid and caffeic acid) also showed the ability to significantly (p<0.001) decrease the parameters analyzed above.
This is the first study to identify and confirm these phenolic compounds in I. asarifolia leaves extract and to suggest that these compounds contribute to the anti-inflammatory activity in vivo, as reported by ethnomedicinal use of this plant. Through the different experimental models performed, we can conclude that the results obtained with the aqueous extract from I. asarifolia leaves support its popular use for the treatment of inflammatory disorders.
圆叶茑萝（Ipomoea asarifolia (Desr.) Roem. and Schult.）（旋花科），俗称萨尔萨或萨尔萨-布拉瓦，是一种植物，其叶煎剂在民间医学中用于治疗各种炎症性疾病，如皮炎、疥疮、梅毒症状、皮肤溃疡和外伤。然而，关于该植物可能的化合物和作用机制，以支持其民间使用所报告的活性，我们知之甚少。
本研究旨在鉴定圆叶茑萝叶粗提物中存在的生物活性分子，并在不同的体内实验模型中研究该提取物的抗炎潜力，以增进对其活性的理解。
通过煎煮（1:10 m/v）制备圆叶茑萝叶水提取物，并通过高效液相色谱-二极管阵列检测器（HPLC-DAD）和液相色谱-二极管阵列检测器-质谱联用（LC-DAD-MS）获得其色谱图。使用以下体内模型评估提取物的潜在抗炎活性：二甲苯诱导的耳水肿（20、30和40mg/kg），评估水肿形成程度；角叉菜胶诱导的腹膜炎（10、20和30mg/kg），在4小时时评估白细胞迁移和细胞因子水平（IL-1β、IL-6、IL-12和TNF-α）；酵母聚糖诱导的气袋炎症（20、30和40mg/kg），在6、24和48小时时通过总计数和分类计数评估白细胞迁移动力学。针对每个实验模型，使用在圆叶茑萝叶水提取物中鉴定出的纯化合物以不同剂量进行相同测试。
通过HPLC-DAD和LC-DAD-MS在圆叶茑萝叶水提取物中鉴定出的化合物为芦丁、绿原酸和咖啡酸。该提取物显著减轻了二甲苯诱导的耳水肿（20、30和40mg/kg剂量分别为81%、85%和86%，p<0.001），以及腹膜炎实验模型中的细胞迁移（10、20和30mg/kg剂量分别为70%、78%和83%，p<0.001）和气袋炎症（20、30和40mg/kg剂量分别为58%、67%和53%，p<0.001）。此外，该提取物显示出显著抑制细胞因子IL-1β、IL-6、IL-12和TNF-α产生的能力（p<0.001）。所测试的次生代谢产物（芦丁、绿原酸和咖啡酸）也显示出显著（p<0.001）降低上述分析参数的能力。
这是首次在圆叶茑萝叶提取物中鉴定并确认这些酚类化合物的研究，并表明这些化合物有助于体内抗炎活性，正如该植物的民族医学用途所报告的那样。通过所进行的不同实验模型，我们可以得出结论，圆叶茑萝叶水提取物获得的结果支持其用于治疗炎症性疾病的民间用途。
