Aqueous Leaf Extract of Jatropha gossypiifolia L. (Euphorbiaceae) Inhibits Enzymatic and Biological Actions of Bothrops jararaca Snake Venom

et al. (2014) Aqueous Leaf Extract of Jatropha gossypiifolia L. (Euphorbiaceae) Inhibits Enzymatic and Biological Actions of Bothrops jararaca Snake Venom. PLoS ONE 9(8): e104952. doi:10.1371/journal.pone.0104952 Aqueous Leaf Extract of Jatropha gossypiifolia L. (Euphorbiaceae) Inhibits Enzymatic and Biological Actions of Bothrops jararaca Snake Venom Juliana Fe lix-Silva 0 Thiago Souza 0 Yamara A. S. Menezes 0 Ba rbara Cabral 0 Rafael B. G. Ca mara 0 Arno bio A. Silva-Junior 0 Hugo A. O. Rocha 0 Ivanise M. M. Rebecchi 0 Silvana M. Zucolotto 0 Matheus F. Fernandes-Pedrosa 0 Luis Eduardo M. Quintas, Universidade Federal do Rio de Janeiro, Brazil 0 1 Laborato rio de Tecnologia & Biotecnologia Farmaceutica (TecBioFar), Programa de Po s-graduac a o em Ciencias Farmaceuticas (PPgCF), Universidade Federal do Rio Grande do Norte (UFRN) , Natal, Rio Grande do Norte, Brazil, 2 Laborato rio de Farmacognosia , Departamento de Farma cia, Universidade Federal do Rio Grande do Norte (UFRN) , Natal, Rio Grande do Norte, Brazil, 3 Laborato rio de Biotecnologia de Pol meros Naturais (BIOPOL) , Programa de Po s-graduac a o em Bioqu mica, Universidade Federal do Rio Grande do Norte (UFRN) , Natal, Rio Grande do Norte, Brazil, 4 Laborato rio de Hematologia Cl nica , Departamento de Ana lises Cl nicas e Toxicolo gicas, Universidade Federal do Rio Grande do Norte (UFRN) , Natal, Rio Grande do Norte , Brazil Snakebites are a serious public health problem due their high morbi-mortality. The main available specific treatment is the antivenom serum therapy, which has some disadvantages, such as poor neutralization of local effects, risk of immunological reactions, high cost and difficult access in some regions. In this context, the search for alternative therapies is relevant. Therefore, the aim of this study was to evaluate the antiophidic properties of Jatropha gossypiifolia, a medicinal plant used in folk medicine to treat snakebites. The aqueous leaf extract of the plant was prepared by decoction and phytochemical analysis revealed the presence of sugars, alkaloids, flavonoids, tannins, terpenes and/or steroids and proteins. The extract was able to inhibit enzymatic and biologic activities induced by Bothrops jararaca snake venom in vitro and in vivo. The blood incoagulability was efficiently inhibited by the extract by oral route. The hemorrhagic and edematogenic local effects were also inhibited, the former by up to 56% and the latter by 100%, in animals treated with extract by oral and intraperitoneal routes, respectively. The inhibition of myotoxic action of B. jararaca reached almost 100%. According to enzymatic tests performed, it is possible to suggest that the antiophidic activity may be due an inhibitory action upon snake venom metalloproteinases (SVMPs) and/or serine proteinases (SVSPs), including fibrinogenolytic enzymes, clotting factors activators and thrombin like enzymes (SVTLEs), as well upon catalytically inactive phospholipases A2 (Lys49 PLA2). Antiinflammatory activity, at least partially, could also be related to the inhibition of local effects. Additionally, protein precipitating and antioxidant activities may also be important features contributing to the activity presented. In conclusion, the results demonstrate the potential antiophidic activity of J. gossypiifolia extract, including its significant action upon local effects, suggesting that it may be used as a new source of bioactive molecules against bothropic venom. - Data Availability: The authors confirm that all data underlying the findings are fully available without restriction. All relevant data are within the paper and its Supporting Information files. Funding: This research was supported by grants from CAPES (23038000814/2011-83), CNPq (483842/2010-9), BNB (ETENE/2010) and FAPERN (PRONEM/2011). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. Snakebites are a serious public health problem in many regions around the world, particularly in tropical and subtropical countries [1,2]. The high morbi-mortality rate still has a great impact on the population and on health-care systems, especially in Africa, Asia, Oceania and Latin America and, unfortunately, public health authorities have given little attention to this problem [1]. Thus, snake envenomation is included in the 2009 World Health Organization (WHO) list of Neglected Tropical Diseases (NTDs) [3]. Conservative estimates indicate that, worldwide, there are more than 5 million snakebites, leading to 25,000125,000 deaths [2,3]. In Brazil, data from Ministry of Health shows that there are more than 25,000 snakebites per year [4]. More than 90% of the snakebites reported every year in Latin America are caused by Bothrops species [5]. In Brazil, the major representatives of the genus are Bothrops jararaca, Bothrops alternatus, Bothrops atrox, Bothrops erythromelas, Bothrops jararacussu and Bothrops moojeni [4]. Despite the existence of evident intraspecific and interspecific variations in the composition and biological activities of their venoms, bothropic venom can induce a qualitatively similar pathophysiological picture, characterized by immediate and prominent local tissue damage (including myonecrosis, hemorrhage and edema), cardiovascular alterations (especially hemorrhage and hypovolemic shock), coagulation disorders (most frequently blood incoagulability) and renal alterations (which could evolve into acute kidney injury) [5]. The snake envenoming is a complex pathophysiological process involving the simultaneous action of different types of toxins, such as snake venom serine proteinases (SVSPs), snake venom metalloproteinases (SVMPs), hyaluronidases and phospholipases A2 (PLA2) [5,6]. Currently, the only available specific treatment is the antivenom serum therapy, which consists of a pool of neutralizing antibodies taken from serum of animals hyperimmunized against toxins of snake venoms. Its effectiveness consists of its ability to provide to the patient antibodies with a high affinity to snake venom, aiming to eliminate the toxins responsible for toxicity of the envenoming [7]. However, the antivenom has some disadvantages, such as limited effectiveness against local effects, risk of immunological reactions (including serum sickness), high cost and difficult access in some regions [5,7]. If antivenom administration is initiated rapidly after envenomation, neutralization of systemic effects is usually achieved successfully. However, neutralization of local tissue damage is more difficult. This poor effectiveness against local effects, as well the increased time between accident and treatment are related to the temporary or permanent disability observed in many victims. It is estimated that 400,000 people are left with permanent disabilities after snakebites [3,5]. In this con (...truncated)