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Anti-snake venom properties of Schizolobium parahyba (Caesalpinoideae) aqueous leaves extract
Abstract
Many medicinal plants have been recommended for the treatment of snakebites. The aqueous extracts prepared from the leaves of Schizolobium parahyba (a plant found in Mata Atlantica in Southeastern Brazil) were assayed for their ability to inhibit some enzymatic and biological activities induced by Bothrops pauloensis and Crotalus durissus terrificus venoms as well as by their isolated toxins neuwiedase (metalloproteinase), BnSP-7 (basic Lys49 PLA(2)) and CB (PLA(2) from crotoxin complex). Phospholipase A(2), coagulant, fibrinogenolytic, hemorrhagic and myotoxic activities induced by B. pauloensis and C. d. terrificus venoms, as well as by their isolated toxins were significantly inhibited when different amounts of S. parahyba were incubated previously with these venoms and toxins before assays. However, when S. parahyba was administered at the same route as the venoms or toxins injections, the tissue local damage, such as hemorrhage and myotoxicity was only partially inhibited. The study also evaluated the inhibitory effect of S. parahyba upon the spreading of venom proteins from the injected area into the systemic circulation. The neutralization of systemic alterations induced by i.m. injection of B. pauloensis venom was evaluated by measuring platelet and plasma fibrinogen levels which were significantly maintained when S. parahyba extract inoculation occurred at the same route after B. pauloensis venom injection. In conclusion, the observations confirmed that the aqueous extract of S. parahyba possesses potent snake venom neutralizing properties. It may be used as an alternative treatment to serum therapy and as a rich source of potential inhibitors of toxins involved in several physiopathological human and animal diseases.
... & Triana (de Oliveira et al., 2014), Bellucia dichotoma Cogn., Aniba fragrans Ducke, Plathymenia foliolosa Benth. (de Moura et al., 2015), Schizolobium parahyba (Mendes et al., 2008), Lapachol (Strauch et al., 2019), Lupeol (Dos Santos et al., 2021). ...
... For in vivo neutralization tests (hemorrhagic and myotoxic activity), two ratios were also used 1:1 (venom/ISO; w/w; e.g. 10 μg Pb + 10 μg ISO) and 1:5 (venom/ISO; w/w; e.g. 10 μg Pb + 50 μg ISO) and two incubation test conditions (ISO) with Pb (10 μg) incubated for 30 min at 37 • C. Treatment application conditions were also performed 15 min after the application of Pb or BthMP (10 μg) and ISO was applied in the same location at the same ratio (1:1 and 1:10, venom/ISO; w/w) according to the methodology used by Mendes and collaborators (Mendes et al., 2008;Mendes et al., 2013). In all the experiments, the final volume was standardized to 50 μL in the tests. ...
... In some studies, it has been suggested that tannins and flavonoids may complex with proteins in a non-specific way, precipitating them (Borges et al., 2005;Mendes et al., 2008;Vale et al., 2011). ISO did not form visible complexes when incubated with proteins, since when the mixture is centrifuged, the presence of precipitate cannot be observed. ...
We investigated the antioph idic properties of isohemigossypolone (ISO), a naph thoquinone isolated from the outer bark of the Pachira aquatic Aubl. The inhibition of ph osph olipase A 2 , coagulant, fibrinogenolytic, hemorrhagic and myotoxic activities indu ced by Bothrops pauloensis venom (Pb) was investigated. For this, we use samples resulting from the incu bation of Pb with ISO in different concentrations (1 :1, 1:5 and 1:10 w/w), we also evaluated the condition of treatme nt using ISO after 15 min of venom inoculation. The activities of ph osph olipase A 2 , coagulant, fibrinogenolytic, hemorrhagic and myotoxic indu ced by the B. pauloen-sis venom were significantly inhibited wh en the ISO was pre-incu bated with the crud e venom. For in vi vo neutralization tests, the results were observed even wh en the ISO was applied after 15 min of inoculation of the venom or me talloprotease (BthMP). Also, to identify the inhibition me ch anism , we perform ed in silico assays, across simulations of molecular coupling and molecular dynamics, it was possible to identify the modes of interaction be tween ISO and bothropic toxins Bm ooMP α-I, Jararacu ssin-I and BNSP-7. The present stud y shows that naph thoquinone isohemigossypolone isolated from the P. aquatica plant inhibited part of the local and systemic damage caused by venom proteins, demonstrating the ph arma cological potential of this compoun d in neutralizing the harm ful effects caused by snakeb ites.
... En muchos países, las plantas han sido utilizadas tradicionalmente en el tratamiento del envenenamiento provocado por serpientes, porque constituyen una excelente fuente de metabolitos farmacológicamente activos, capaces de antagonizar los lesivos efectos del veneno (15), convirtiéndose en una promisoria alternativa terapéutica para mejorar el pronóstico y evolución de los pacientes tratados con antiveneno, cuya utilización acarrea problemas de hipersensibilidad y limitada protección contra Figura 2. Porcentaje de inhibición de la actividad hemolítica indirecta de las fracciones obtenidas de planta completa de Renealmia alpinia (Rottb) Mass, obtenida en medios de micropropagación. los efectos locales de rápida instalación, y cuyo almacenamiento y administración requiere exigentes condiciones (26)(27)(28)(29). Se ha informado sobre un gran número de plantas con significativas propiedades antiofídicas de uso tradicional en Colombia. ...
... Concretamente se ha informado sobre propiedades antiofídicas de varias cumarinas como la umbeliferona, que interacciona con la PLA 2 del veneno de Crotalus durissus collilineatus , ocasionándole fuertes modificaciones estructurales que se traducen en la supresión del fenómeno inflamatorio producido por la toxina (31). Aunque no se conoce el mecanismo de acción, muchos autores sugieren que tal efecto inhibitorio es promovido por unión de estos compuestos a iones divalentes como el Ca 2+ y el Zn 2+ , importantes para la actividad enzimática de PLA 2 y metaloproteinasas, o a interacciones entre los compuestos y las toxinas, que pueden promover pérdida de la actividad biológica de éstas (28,32). Sin embargo, es necesario un nuevo estudio que permita caracterizar cada una de las posibles cumarinas aisladas y comprobar si su posible efecto quelante, capaz de inhibir la actividad de PLA 2 , es reversible o irreversible cuando se adiciona un exceso de iones de Ca 2+ , como ocurre con los extractos acuosos de Musa paradisiaca y hojas de Schizolobium parahyba (28,33). ...
... Aunque no se conoce el mecanismo de acción, muchos autores sugieren que tal efecto inhibitorio es promovido por unión de estos compuestos a iones divalentes como el Ca 2+ y el Zn 2+ , importantes para la actividad enzimática de PLA 2 y metaloproteinasas, o a interacciones entre los compuestos y las toxinas, que pueden promover pérdida de la actividad biológica de éstas (28,32). Sin embargo, es necesario un nuevo estudio que permita caracterizar cada una de las posibles cumarinas aisladas y comprobar si su posible efecto quelante, capaz de inhibir la actividad de PLA 2 , es reversible o irreversible cuando se adiciona un exceso de iones de Ca 2+ , como ocurre con los extractos acuosos de Musa paradisiaca y hojas de Schizolobium parahyba (28,33). ...
Plant extracts are a rich source of pharmacologically active molecules. Their application allows a tradicional medicine approach to potential biological acitivities. This paper evaluates the inhibitory capacity of ethanolic extracts of leaves and little roots and also fractions chromatographically fractions obtained from Renealmia alpinia (Rottb) Mass, cultured in vitro, on the indirect hemolytic activity, proteolytic activity and coagulant activity induced by the Bothrops asper venom. Indirect hemolytic activity is inhibited to a greater extent by the fraction 7-8 (47.3 +/- 2.20%) followed in order by the extracts from little roots (32.6 +/- 6.90%) and leaves (24.2 +/- 4.43%). They came from in vitro and ex vitro leaves (16.2 +/- 3.88%). The proteolytic activity is largely inhibited by the leaves extracts in vitro and ex vitro without significant differences between them. Little roots in vitro showed the highest neutralization effect on coagulant activity (81.73 +/- 9.949s). Proteolytic activity from Renealmia alpinia extracts on Bothrops asper venom is ruled out since there are not changes in the electrophoretic pattern of the venom. The results make possible the implementation of Renealmia alpinia as adjuvant for the treatment of ophidic accidents and sustain the value of micropropagation for mass production of active components.
... It grows in tropical forests and has been planted extensively in reforestation projects from the Amazon Basin south to São Paulo [36]. SP is popularly known as faveira, guapuruvú or umbela, and healers and shamans have traditionally used SP infusions as natural medicines for treating snakebite envenoming [37][38][39]. In Brazil, healers traditionally use leaves from the Schizolobium parahyba tree to treat venomous snakebite accidents. ...
... In Brazil, healers traditionally use leaves from the Schizolobium parahyba tree to treat venomous snakebite accidents. Experimental studies have shown that gross aqueous extract and flavonoids from Schizolobium parahyba protect against some of the effects of Bothrops venom in rodent animal models and in vitro preparations [37][38][39]. ...
... On the experiment day, SP was dissolved in 10 ml of 0.9% saline and infused via the jugular vein at a dose of 2.0 mg/kg at a rate of 0.02 ml/min for 40 min (Compact Infusion Pump, 975, Harvard, USA). This dose was selected from previous publications (37)(38)(39). ...
Venom-induced acute kidney injury (AKI) is a frequent complication of Bothrops snakebite with relevant morbidity and mortality. The aim of this study was to assess the effects of Schizolobium parahyba (SP) extract, a natural medicine with presumed anti-Bothrops venom effects, in an experimental model of Bothrops jararaca venom (BV)-induced AKI.
Groups of 8 to 10 rats received infusions of 0.9% saline (control, C), SP 2 mg/kg, BV 0.25 mg/kg and BV immediately followed by SP (treatment, T) in the doses already described. After the respective infusions, animals were assessed for their glomerular filtration rate (GFR, inulin clearance), renal blood flow (RBF, Doppler), blood pressure (BP, intra-arterial transducer), renal vascular resistance (RVR), urinary osmolality (UO, freezing point), urinary neutrophil gelatinase-associated lipocalin (NGAL, enzyme-linked immunosorbent assay [ELISA]), lactate dehydrogenase (LDH, kinetic method), hematocrit (Hct, microhematocrit), fibrinogen (Fi, Klauss modified) and blinded renal histology (acute tubular necrosis score).
BV caused significant decreases in GFR, RBF, UO, HcT and Fi; significant increases in RVR, NGAL and LDH; and acute tubular necrosis. SP did not prevent these changes; instead, it caused a significant decrease in GFR when used alone.
SP administered simultaneously with BV, in an approximate 10∶1 concentration, did not prevent BV-induced AKI, hemolysis and fibrinogen consumption. SP used alone caused a decrease in GFR.
... The aqueous extract of leaves of Schizolobium parahyba significantly inhibited the coagulant, hemorrhagic and fibrinogenolytic activities induced by Bothrops pauloensis and Crotalus durissus terrificcus venoms and their isolated toxins after preincubation with venoms and toxins before assays.[29] In vivo tests with polyphenols of Areca catechu L and Quercus infectoria Oliv showed inhibition of the hemorrhagic activity of Calloselasma rhodostoma Kuhl venom and dermonecrotic activity of Naja kauothia venom.[30] ...
... Neutralization of myotoxic effects of Vipera russelii venom is reported with the bark extract of Anacardium occidentale.[20] Significant inhibition of myotoxicity induced by Bothrops pauloensis and Crotalus durissus terrificcus venoms and their isolated toxins by aqueous extract of leaves of Schizolobium parahyba has been documented.[29] ...
... Inhibition of enzymatic activity is reported with extracts of Casearia sylvestris in experimental animals, injected with lethal doses of Bothropic venoms.[16] Significant inhibition of PLA2 activity induced by Bothrops pauloensis and Crotalus durissus
terrificcus venoms is documented with the leaf extract of Schizolobium parahyba.[29] Neutralization of Vipera russelii venom enzymes namely phospholipase, protease and hyaluronidase is reported with the bark extract of Anacardium occidentale in a dose-dependent manner.[20] ...
Snake envenomation is a global public health problem, with highest incidence in Southeast Asia. Inadequate health services, difficult transportation and consequent delay in antisnake venom administration are the main reasons for high mortality. Adverse drug reactions and inadequate storage conditions limit the use of antisnake venom. The medicinal plants, available locally and used widely by traditional healers, therefore need attention. A wide array of plants and their active principles have been evaluated for pharmacological properties. However, numerous unexplored plants claimed to be antidotes in folklore medicine need to be studied. The present article reviews the current status of various medicinal plants for the management of snake bite.
... Tamarindus indica seed extract has stopped the hemorrhage, indirect hemolysis and degradation of beta chain of human fibrinogen, caused by viper venom in experimental animals [39] . The aqueous extract of leaves of Schizolobium parahyba notably inhibited the coagulant, hemorrhagic and fibrinogenolytic properties of Bothrops pauloensis and Crotalus durissus terrificcus venoms and their isolated toxins after pre-incubation with venoms and toxins before assays [45] . ...
... Inhibition of enzymatic activity is pronounced with extracts of Casearia sylvestris in experimental animals, injected with deadly doses of Bothropic venoms [36]. Substantial inhibition of PLA2 activity prompted with the aid of Bothrops pauloensis and Crotalus durissus terrificcus venoms is documented with the leaf extract of Schizolobium parahyba [45] . Neutralization of Vipera russelii venom enzymes specifically phospholipase, protease and hyaluronidase is reported with the bark extract of Anacardium occidentale in a dose-based way [40] . ...
The mortality related to snake bites is an extreme public health problem because the estimated death prevalence per year is about 1,25,000 globally. Insufficient health services, bad transportation and consequent delay in synthetic anti-snake venom management are the main motives for excessive mortality. Adverse drug reactions and inadequate storage conditions limit the use of synthetic anti-snake venom. The medicinal flora, available domestically and used widely by traditional healers, consequently need attention. Most of the snake bite cases are typically undermined due to loss of proper consciousness of the mass people. However, present evaluation has been focused at the artificial and traditional herbs and their anti-venom property, which can be a venturing stone in setting up the future treatment against snake bite treatment and management.
... Tamarindus indica seed extract has stopped the hemorrhage, indirect hemolysis and degradation of beta chain of human fibrinogen, caused by viper venom in experimental animals [39] . The aqueous extract of leaves of Schizolobium parahyba notably inhibited the coagulant, hemorrhagic and fibrinogenolytic properties of Bothrops pauloensis and Crotalus durissus terrificcus venoms and their isolated toxins after pre-incubation with venoms and toxins before assays [45] . ...
... Inhibition of enzymatic activity is pronounced with extracts of Casearia sylvestris in experimental animals, injected with deadly doses of Bothropic venoms [36]. Substantial inhibition of PLA2 activity prompted with the aid of Bothrops pauloensis and Crotalus durissus terrificcus venoms is documented with the leaf extract of Schizolobium parahyba [45] . Neutralization of Vipera russelii venom enzymes specifically phospholipase, protease and hyaluronidase is reported with the bark extract of Anacardium occidentale in a dose-based way [40] . ...
The mortality related to snake bites is an extreme public health problem because the estimated death prevalence per year is about 1,25,000 globally. Insufficient health services, bad transportation and consequent delay in synthetic anti-snake venom management are the main motives for excessive mortality. Adverse drug reactions and inadequate storage conditions limit the use of synthetic anti-snake venom. The medicinal flora, available domestically and used widely by traditional healers, consequently need attention. Most of the snake bite cases are typically undermined due to loss of proper consciousness of the mass people. However, present evaluation has been focused at the artificial and traditional herbs and their anti-venom property, which can be a venturing stone in setting up the future treatment against snake bite treatment and management.
... Although their use for the treatment of snakebites is traditional in many countries, plant extracts have been shown empirically to be a promising alternative for this purpose, but without scientific evidence of their efficacy [13]. Several studies are speculating on the use of plant extracts as sources of molecular prototypes which can be used in the treatment of snake envenomations, or to complement the traditional antivenoms, which have shown little effectiveness in minimizing the local damage [3,[13][14][15][16][17][18][19]. However, studies employing essential oils for that purpose are still scarce [18]. ...
... Fibrinogen, a dimeric plasma glycoprotein of 340 kDa composed of six polypeptide chains (α, β, γ), participates in the coagulation cascade, and is converted by thrombin to fibrin monomers [37]. This protein has been extensively used for the evaluation of the potential fibrinogenolytic effects of snake venoms, and for the verification of potential antiophidian properties of natural compounds, mainly of plant origin [3,13,[15][16][17][18]38,39]. ...
The search for new inhibitors of snake venom toxins is essential to complement or even replace traditional antivenom therapy, especially in relation to compounds that neutralize the local effects of envenomations. Besides their possible use as alternative to traditional antivenom therapy, some plant species possess bioactive secondary metabolites including essential oils, which can be extracted from weeds that are considered substantial problems for agriculture, such as Hedychium coronarium.
The essential oils of leaves and rhizomes from H. coronarium were extracted by hydrodistillation, and their potential inhibitory effects on the coagulant and fibrinogenolytic activities induced by the venoms of Lachesis muta, Bothrops atrox and Bothrops moojeni were analyzed. Citrated human plasma was used to evaluate the clotting time whereas changes in fibrinogen molecules were visualized by electrophoresis in polyacrylamide gel. The experimental design used for testing coagulation inhibition was randomized in a 3 × 2 factorial arrangement (concentration × essential oils), with three replications. The essential oils were compared since they were extracted from different organs of the same botanical species, H. coronarium.
The results suggest that the oils interact with venom proteases and plasma constituents, since all oils evaluated, when previously incubated with venoms, were able to inhibit the clotting effect, with less inhibition when oils and plasma were preincubated prior to the addition of venoms.
Thus, after extensive characterization of their pharmacological and toxicological effects, the essential oils can be used as an alternative to complement serum therapy, especially considering that these plant metabolites generally do not require specific formulations and may be used topically immediately after extraction.
... As for leukocytes (Table 2), in our model no significant changes were found in the experimental groups. On the other hand, only B. asper venom significantly diminished the number of platelets, a finding similar to what Mendes et al. (18) reported in mice and in other human case reports (19). The current contribution shows that Randia aculeata protects against this event, which is why we infer that one of the plant's effects could be avoiding hemorrhages otherwise caused by snake venom, as has been demonstrated by employing other vegetal species (20)(21)(22)(23). ...
... The current contribution shows that Randia aculeata protects against this event, which is why we infer that one of the plant's effects could be avoiding hemorrhages otherwise caused by snake venom, as has been demonstrated by employing other vegetal species (20)(21)(22)(23). It is also possible that this plant can partially neutralize myotoxic effects caused by some snake venoms, which are caused by phospholipase A 2 , metalloproteases and crotoxins (18,24). It has been proven that some plants can neutralize these toxins in vitro (23,25). ...
In Mexico, medicinal plants are widely used. The use of Randia aculeata by healers against snakebites has never been scientifically tested in relation to possible effects on blood parameters and muscle tissue damage. Interviews were carried out in Jamapa, Veracuz, Mexico, with local residents to collect information about the traditional use of Randia aculeata. In this locality, seven pieces of fruit from the plant are mixed in a liter of alcohol, and then administered orally against snakebites. By using histological techniques and a murine model, we explored its cytoprotective properties against the effects of Crotalus simus and Bothrops asper venoms. Possible protections provided by the plant against tissue damage to skeletal and cardiac muscles and against the typical loss of red blood cells were analyzed. Randia aculeata caused an increase in microhematocrit and total hemoglobin, parameters that are often decremented in association with the loss of red blood cells, which is a characteristic effect of animal venom. Randia aculeata was also shown to protect against the lowering of platelet levels caused by Bothrops asper venom. Finally, Randia aculeata produced a partial inhibition of necrosis following administration of snake venom in skeletal and myocardial muscles. The present results provide solid evidence for the traditional use of Randia aculeata against snakebites, as demonstrated by protection against muscular tissue damage and the diminution of red blood cells.
... Several vegetal extracts contain compounds capable of neutralizing the pro-coagulant and anticoagulant activities of snake venoms (Borges et al., 2000(Borges et al., , 2005Biondo et al., 2003;Mendes et al., 2008;Vale et al., 2011). In this work, the inhibition of coagulant activity on bovine plasma induced by the B. pauloensis snake venom by the Bg extract and PCT was demonstrated. ...
... Histological cuts of 5 lm were made and stained with hematoxylin and eosin (HE) for light microscopy and morphological alteration evaluation. Control mice received only saline or PCT (4) (Rodrigues et al., 2001;Mendes et al., 2008Mendes et al., , 2010. ...
Snake venom metalloproteinases (SVMPs) participate in a number of important biological, physiological and pathophysiological processes and are primarily responsible for the local tissue damage characteristic of viperid snake envenomations. The use of medicinal plant extracts as antidotes against animal venoms is an old practice, especially against snake envenomations. Such plants are sources of many pharmacologically active compounds and have been shown to antagonize the effects of some venoms and toxins. The present study explores the activity of triacontyl p-coumarate (PCT (4)), an active compound isolated from root bark of Bombacopsis glabra vegetal extract (Bg), against harmful effects of Bothropoides pauloensis snake venom and isolated toxins (SVMPs or phospholipase A(2)). Before inhibition assays, Bg or PCT (4) was incubated with venom or toxins at ratios of 1:1 and 1:5 (w/w; venom or isolated toxins/PCT (4)) for 30min at 37°C. Treatment conditions were also assayed to simulate snakebite with PCT (4) inoculated at either the same venom or toxin site. PCT (4) neutralized fibrinogenolytic activity and plasmatic fibrinogen depletion induced by B. pauloensis venom or isolated toxin. PCT (4) also efficiently inhibited the hemorrhagic (3MDH - minimum hemorrhagic dose injected i.d into mice) and myotoxic activities induced by Jararhagin, a metalloproteinase from B. jararaca at 1:5 ratio (toxin: inhibitor, w/w) when it was previously incubated with PCT (4) and injected into mice or when PCT (4) was administered after toxin injection. Docking simulations using data on a metalloproteinase (Neuwiedase) structure suggest that the binding between the protein and the inhibitor occurs mainly in the active site region causing blockade of the enzymatic reaction by displacement of catalytic water. Steric hindrance may also play a role in the mechanism since the PCT (4) hydrophobic tail was found to interact with the loop associated with substrate anchorage. Thus, PCT (4) may provide a alternative to complement ophidian envenomation treatments.
... Brazilian researchers have recently assessed the effects of a regional popular medicine against snake venom which is commonly used in the central region to treat snakebites [58,59] . They evaluated the effect of the administration of Schizolobium parahyba (Fabaceae) infusion and its fractions against the pharmacological and toxic activity of Bothrops venom in rats. ...
... This native plant from the Brazilian forest, popularly known as guapuruvú, was effective in protecting against some of the B. alternatus and B. moojeni venom-induced enzymatic and biological activities. Animal lethality was reduced to zero and the venom hemolytic, hemorrhagic and coagulation system effects were attenuated [58,59] . Studies aiming at Brazilian forest phytotherapy are a sustainable research choice. ...
Medically important venomous snakes in Latin America belong to the genus Bothrops, Crotalus, Lachesis and Micrurus. The Bothrops genus is responsible for the majority of accidents. The WHO globally estimates 2,500,000 poisonous snakebites and 125,000 deaths annually. In its last report in 2001, the Brazilian Ministry of Health accounted 359 deaths due to snakebites, of which the Bothrops genus was responsible for 185. Snake venoms cause local and systemic damage, including acute kidney injury, which is the most important cause of death among patients surviving the early effects of envenoming by the Crotalus and Bothrops genuses. Venom-induced acute kidney injury is a frequent complication of Bothrops snakebite, carrying relevant morbidity and mortality.
... Schizolobium parahyba, popularly known as 'faveira, guapuruvu or umbela', is a plant used in popular medicine to treat snakebites in the Brazil central region, called 'Triângulo Mineiro'. In recent studies, the herbal extract of S. parahyba leaves and fractions was shown to possess potent secondary metabolites that are able to inhibit local tissue damage, such as hemorrhage and myotoxicity induced by Bothrops snake venoms (Vale et al., 2007;Mendes et al., 2008). Other studies with this plant showed the isolation of serine proteinase inhibitors from its seeds (Mizuta and Ventura, 1976;Souza et al., 1995;Sampaio et al., 1996;Teles et al., 2004). ...
... Herbal extracts present a wide range of antiophidian activities, however, in most cases, scientifi c evidence of these activities is still necessary. Several plants have already been proved to display antivenom activity (Martz, 1992;Mors et al., 2000;Soares et al., 2004Soares et al., , 2005Vale et al. 2007;Mendes et al., 2008). ...
The herbal extract of Schizolobium parahyba leaves is used commonly in the Brazil central region to treat snakebites. This study evaluates the acute toxicological effects of Schizolobium parahyba aqueous extract in mice 24 h after intraperitoneal administration. Acute toxicity was evaluated using biochemical, hematological and histopathological assays. Alterations in the levels of transaminases, bilirubin, albumin and prothrombrin time were observed, and these are likely to occur due to hepatic injury, which was confirmed by light microscopy. Liver histopathological analysis revealed the presence of lymph plasmocitary inflammatory infiltrate, but no other histopathological alterations were observed in any of the other organs analysed. The data confirm the low toxicity of the extract of Schizolobium parahyba and provide a model for the selection of a dose that does not cause injuries in the organism.
... Guapuruvu, "tronco de fazer canoa": Schizolbium parahyba (Vell) Bignoniaceae Conhecimento popular: as folhas contêm substâncias que atuam como antídoto para as picadas de jararaca. Academia: observações confirmaram que o extrato aquoso de S. parahyba tem propriedades potentes de neutralização do veneno da serpente (MENDES et al., 2008). ...
... In this respect, several plants have been studied for their potential antiophidic activity, including Schizolobium parahyba (Mendes et al., 2008, Bombacopsis glabra (Mendes et al., 2012), Bellucia dichotoma, Connarus favosus (Moura et al., 2015), Tamarindus indica, Paullinia pinnata (Molander et al., 2015), Bredemeyera floribunda , Zanthoxylum monogynum and Paquira aquática (Vieira et al., 2021). ...
... Also, there are plants that exhibit anti-ophidic effects such as the one mentioned earlier Casearia decandra Jacq, and the Schizolobium parahyba (Vell) species of the family Caesalpiniaceae, S.F. Blake, popularly known as guapuruvu, whose leaves can be used to neutralize intoxication by snakes (Mendes, et al., 2008). Other plants that have effects against external pathogens and were already mention in here are Allophylus edulis Radl, Psidium cattleianum Sabine. ...
The Atlantic Forest Biome is essential for 72% of the Brazilian population, but only a small part of the population has knowledge and values this environmental heritage. This biome is under constant threat; thus, strategies and actions are needed for its preservation and conservation. The Embu-Verde Environmental Preservation Area (EPA) concentrates a big part of the remaining biodiversity of the Atlantic Forest, and because it is close to the largest urban center in the country, it is increasingly threatened. The use of plants as a medicinal tool has been explored since the beginning of mankind. In fact, many of the compounds used in the health area derive from the flora. In this context, this study aimed to identify the medicinal potential of the EPA Embu-Verde flora, from the perspective that it is necessary to know to be able to preserve. Based on previous studies registering the biodiversity of EPA Embu-Verde, all the species of flora listed were investigated to identify its medicinal potential. We show here that of the 202 native species identified in the EPA Embu-Verde, there are 35 species of 23 botanical families that presented medicinal potential. The Myrtaceae family was the most cited, and its anti-inflammatory potential was the most prominent, moreover, several other uses were reported for the remaining flora. The medicinal potential of the flora of EPA Embu-Verde is still underestimated, therefore the recognition of medicinal species by the population is important. The preservation of the flora is essential to guarantee the possibility of future bioprospecting studies that will investigate the therapeutic potential of the still unknown medicinal species.
... The most widely used anticoagulant is heparin. However, because it has animal origin, it can induce diseases in mammals such as avian influenza and bovine spongiform encephalopathy (Mendes et al., 2008), reinforcing the need to find new anticoagulants and antithrombotic agents from plants. ...
Snake venom are widely used as laboratory tools for studies of physiological, pharmacological and toxicological mechanisms. Venoms used here are rich sources of several classes of proteases that act on factors of the coagulation cascade, fibrinogenolysis and fibrinolysis, altering the hemostatic processes, and phospholipases A2 which are involved mainly in inflammatory and clotting processes since they act hydrolyzing membrane phospholipids and may result in the release of arachidonic acid whose structure is a precursor of eicosanoids by cyclooxygenase and lipoxygenase pathways. Natural products such as essential oils are made up of active ingredients with wide application in the food, pharmaceutical and cosmetic industries. Thus, in this study evaluate the essential oils from Mentha viridis and Mentha pulegium on coagulation, fibrinogenolysis and degradation of azocasein, induced by Bothrops sp and Lachesis muta muta venoms. These oils were achieved by hydrodistillation and presented, respectively, as the main constituents linalool (40.70%), carvone (13.52%) and α-terpinene (8.56%); pulegone (50.01%), menthol (31.90%) and menthone (16.56%). The essential oils were previously incubated with Bothrops alternatus venom, for two different times, plasma was added and timing. The M. pulegium and M. viridis oils in the volume of 0.30 μL (10 min of incubation) presented greater anticlotting potential. Meanwhile, 0.15 μL the M. pulegium oil presented proclotting activity. In 20 min of incubation, both oils presented anticlotting activity with 0.15 and 0.30 μL. At azocaseinolytic assay the oil from M. pulegium reduced the activity for all evaluated venoms. The highest inhibitions were 34.33% and 39.99% for 0.6 and 1.2 μL of oil, respectively; on activity induced by B. jararacussu, M. viridis with 0.6 and 1.2 μL reduced the activity in 40.93% and 57.72%, respectively. On B. moojeni, the same volumes were responsible for inhibitions of 74.67% and 47.4%, respectively. The fibrinogenolysis induced by B. moojeni venom was totally inhibited by both oils in the evaluated proportions. The results show the presence in oils of protease inhibitors, considering metalloproteases (mainly with thrombin-like and hemorrhagic activity) and serineproteases (actuating on clotting factors), as well as phospholipase A2 inhibitors (involved in inflammation and clotting processes) of wide application in medical and biotechnology areas.
... This plant is commonly found in the Atlantic rainforest in southeastern Brazil and is popularly known as faveira, guapuruvú, or umbela. It is used as infusions against snakebites and the studies have shown that its leaf extract decreased the hemorrhage and myotoxicity when venom and the extract were previously incubated or the extract was administered within 15 min of venom injection by the same route [75,76]. Another study showed that four flavonoids isolated from this plant including, isoquercitrin 13, catechin 14, myricetin-3-O-glucoside 15, and gallocatechin 16, were able to inhibit the hemorrhagic and fibrinogenolytic activities of SVMP from B. jararacussu and B. neuwiedi venoms. ...
Snakebite envenomation is an important health problem in tropical countries, with severe human and social consequences. In Latin America, the Bothrops species constitutes the main threat to humans, and the envenomation caused by these species quickly develop into severe local tissue damage, including swelling, hemorrhaging, myonecrosis, skin ulceration, and pain. The systemic effects of envenomation are usually neutralized by antivenom serum therapy, despite its intrinsic risks. However, neutralization of local tissue damage remains a challenge. To improve actual therapy, two major alternatives are proposed: the rational design of new specific antibodies for most of the tissue damaging/poor immunogenic toxins; or the search for new synthetic or natural compounds able to inhibit these toxins and complement the serum therapy. Natural compounds isolated from plants, mainly from those used in folk medicine to treat snakebite, are a good choice for finding new lead compounds to improve snakebite treatment and minimize its consequences for the victims. In this article, we reviewed the most promising plants and phytocompounds active against bothropic venoms.
... Numerous attempts have been made by researchers to develop snake venom antagonists from plants on the basis of the fact that these medicinal plants possess rich bioactive compound with potent pharmacological acti- vity (Martz, 1992;Soares et al., 2005). The plants are Eclipta sp., Casearia sp., Curcuma longa, Mimosa pudica ( Meenatchisundaram et al., 2009), Musa paradisiaca, Mucuna pruriens, Bauhinia forficata, Hibiscus esculentus, Annoma senegallensis, Mikania glomerate ( Floriano et al., 2009), Piper sp, Schizolobium parahyba ( Mendes et al., 2008) etc. ...
p class="Abstract">Anti-snake venom therapy is the only treatment for snake bite but leads to acute and chronic conditions which may be severe. The medicinal plants have gained importance over years to find an effective alternative to anti-snake venom. The present study focused on evaluating the potential of Clerodendrum serratum for the anti-snake venom activity. Phytochemicals were extracted from the C. serratum with different solvents. The ethyl acetate and methanolic extracts were found to neutralize the major enzyme toxins (phospholipase A<sub>2</sub>, protease and hyaluronidase) of Bungarus caeruleus and Daboia russelii venom at a concentration of 100 µg/mL. The fibrinogenolytic activity of both the venoms were neutralized. The study proves that the plant C. serratum possesses certain compounds which inhibit the toxins present in the venom of B. caeruleus and D. russelii .
Video Clip of Methodology :
Hyaluronidase assay: 3 min 30 sec Full Screen Alternate </p
... Studies have shown that guapuruvu wood is more susceptible to color change than other species (Mattos et al., 2016). There are also studies about the medicinal uses of extracts from its leaves (Vale et al., 2011;Mendes et al., 2008). ...
Guapuruvu is one of the fastest growing Atlantic Forest species. It is also considered one of most important species in forest recovery areas, starting to compete with other individuals during the second stage of succession, as well as showing a capacity to be harvested and processed at this stage, after fulfilling its function as a pioneer. There are no conclusive studies about the technological potential of the wood from the young guapuruvu trees characteristic of this context. The present study evaluated the physical-mechanical properties and quality of guapuruvu wood from young trees (15 years old) from forest recovery areas in accordance with NBR 7190. The presence of tension wood did not influence the homogeneity of the wood, which is classified as strength class C20 – hardwood, with characteristic strength of 21.47 MPa in compression parallel to the grain, characteristic tension strength of 23.12 MPa parallel to grain and modulus of elasticity (MOE) of 5,925 MPa. Basic and apparent densities were 290 kg/m3 and 336 kg/m3 respectively. However, its low density did not lead to low mechanical strength. The high ratio between the values of its mechanical properties and apparent density, added to its high homogeneity – despite the presence of tension wood – demonstrated the good quality of its wood.
... The activity of plasmatic creatine kinase was determined with the aid of a commercial kit (Labtest, Brazil). The activity was expressed in units/L, with one unit corresponding to the production of 1mmol of NADH per min at 30 C [23]. The anti-myotoxic potential of antibodies was determined as percent inhibition of myotoxic activity. ...
Polyclonal antibodies raised in Balb-c mice against BnSP-7, a Lys-49 phospholipase A2, were used to measure cross reactivity against other snake venoms. Using ELISA, these antibodies were able to recognize PLA2s isoforms present in venoms of botropic snakes at 1:6400, 1:3200 and 1:100 ratios (w/w). These antibodies highly recognized proteins of low molecular weight (∼14,000) from crude snake venom Bp and Bm by Western Blotting. PLA2 these venoms, by alignment of primary structures demonstrated high identity with BnSP-7 PLA2, especially in the C-terminal region. However, the crude snake venom Bd and Bj, showed low recognition. The PLA2 activity of Bothrops pauloensis, Bothrops moojeni venoms or BpPLA2-TXI was inhibited significantly when anti-BnSP-7 antibodies were incubated at 1:10 and 1:20 ratios (venoms or toxin:anti-BnSP-7, w/w), respectively. The myotoxic effect induced by the same venoms was also reduced significantly at 1:1, 1:10 and 1:20 ratios, by decreased creatine kinase levels. The anti-PLA2 polyclonal antibodies effectively recognized PLA2s from Bothrops pauloensis and Bothrops moojeni venoms, and neutralized specific catalytic and myotoxic activity.
... Enzymatic activity was expressed as percentage and inhibition of enzymatic activity was recorded using pre-incubated venom with different plant extracts. Inhibition of enzymatic activity was measured by pre-treated venom (5 μg) with different plant extracts at a concentration of 5 μg/ml for 30 min 15,16 . ...
Phospholipase A (PLA) is ubiquitous in nature and the most noxious component of Naja naja karachiensis venom. The present work was designed to study the role of PLA as anticoagulant in an egg yolk assay and to assess the potential of the extracts of 29 medicinal plants of Pakistan to counter this action. Coagulation assays with venom were performed in vitro with hen's egg yolk mixture. Subsequently this effect was endeavoured to neutralize with plants extract and compared with standard antisera. Venom at concentration of 5 μg clotted within 125 sec compared to venom-free egg yolk (control) mixture which clotted in 100 sec. Citrullus colocynthis, Rubia cordifolia and Stenolobium stans (11% extracts) were found to stop anticoagulant action as recorded with reference standard antidote. On the other hand, Albizia lebbeck, Brassica nigra, Matthiloa incana, Nerium indicum and Rhazya stricta (17% extracts) were declared completely abortive. Moreover, Allium cepa, Allium sativum, Althaea officinalis, Bauhinia variegate, Calotropis procera (flowers), Calotropis procera (exudates), Cedrus deodara, Citrus limon, Cuminum cyminum, Enicostemma hyssopifolium, Fogonia cretica, Leucas capitata, Momordica charantia, Ocimum sanctum, Pinus roxburghii, Pistacia integerrima, Psoralea corylifolia, Sapindus mukorossi, Terminalia arjuna, Trichodesma indicum and Zingiber officinalis (72% extracts) showed restoration against anticoagulation behaviour, i.e. anticoagulation decreased from 92% to 20%. However, further research is inevitable for isolation and structure elucidation of bioactive constituent(s) from active plants extract.
... A quantidade de acidentes ofídicos notificados no Brasil central, apesar de baixa é considerada como um problema de saúde pública, pois quando o paciente é tratado com soro antiofídico de forma efetiva, os efeitos sistêmicos são neutralizados, enquanto os locais não são revertidos, havendo uma possibilidade real de surgirem sequelas (ANAI et al, 2002). Diante disso, muitos pesquisadores vêm buscando as plantas medicinais como terapia alternativa, devido ao seu papel antiofídico (IZIDORO et al., 2003;SOARES et al., 2005;VALE et al., 2008;MENDES et al., 2008;MENDES et al., 2010). Segundo Matsuda et al. (2002) e Chimnoi et al. (2009) da planta Hedychium coronarium já foi isolado alguns tipos de diterpenos que baseado na literatura, estas moléculas são caracterizadas como antiofídicas (MORS et al., 2004). ...
Envenenamentos com serpentes do gênero Bothrops podem causar sequelas no local da picada, que não são revertidas mesmo após o tratamento com soro antiofídico. A incubação do extrato aquoso de Hedychium coronarium (Zingeberaceae) com veneno da serpente Bothrops pauloensis em diferentes concentrações foi capaz de inibir algumas atividades enzimáticas. No presente trabalho ajustou-se um modelo de regressão entre níveis de concentração de extrato e tempo de coagulação (segundos). O modelo ajustado conseguiu captar cerca 96 % da variação total do tempo de coagulação.
... Snake bites pose a major health risk in many countries, with the global incidence of snake bites exceeding 5,000,000 per year (Williams et al., 2010). This problem is more profound in the developing countries, particularly in areas where the access to medical service and to the antiophidic treatment is challenging (Mendes et al., 2008). Although, the majority of snake species are non-venomous and typically kill their prey with constriction, venomous snakes can be found on every continent except Antarctica (Kasturiratne et al., 2008). ...
Olax viridis (Olacaceae) and Syzygium guineense (Myrtaceae) are shrubs commonly found in the tropics. They are traditional folkloric medicine for a great number of sicknesses. Olax viridis has a wide range of applications in ethnomedicine which include treatment for ulcers, veneral diseases, ringworm, sleeping sickness, diarrhea, fever. Syzygium guineense has been reported as antidiarrheal agent. Liquid from the bark and roots have been reported to act as a purgative when mixed with water. Both plants have been claimed to have antivenom properties. However, there are no scientific reports on snake venom neutralizing activities of these plants. The plant samples were collected from Olowa in Dekina Local Government Area in Kogi State, Nigeria. The chemicals and reagents used were of analytical grade. Wistar albino rats (male) weighing between 180-200 g were randomly divided into seven groups of three (3). Groups 1-7 received water, normal saline, venom, venom and Olax viridis, venom and Syzygium guineense, Olax viridis and Syzygium guineense respectively. The extracts were administered orally at the dose of 400 mg kg −1 b.w of rats and 1 h later, the venom (0.08 mk kg −1) was administered. Pulse rate, blood glucose, rectal temperature, plasma cholesterol, triacylglycerol, creatine kinase activity and edema were measured. Significant neutralization of the effects of Naja katiensis venom was observed in the groups of rats that received the extracts. Blood glucose, pulse rate, rectal temperature and creatine kinase activity were elevated in the untreated envenomated groups. These results suggest that oral administration of Olax viridis and Syzygium guineense extracts possess antivenom property, thus, providing the rationale for their use in treatment of sake envenomation.
... Mendes and collaborators [46] reported that the aqueous extract of Schizolobium parahyba (Fabaceae), a plant found in the Mata Atlântica of southeastern Brazil, contains compounds that can inhibit some enzymatic and biological activities induced by Bothrops pauloensis (current Bothropoides paulensis) and C. d. terrificus snake venom as well as by their isolated neuwiedase toxins (metalloproteinase), BnSP-7 (basic Lys49-PLA 2 from B. paulensis venom), and Crotoxin B. ...
Ophidian envenomation is an important health problem in Brazil and other South American countries. In folk medicine, especially in developing countries, several vegetal species are employed for the treatment of snakebites in communities that lack prompt access to serum therapy. However, the identification and characterization of the effects of several new plants or their isolated compounds, which are able to inhibit the activities of snake venom, are extremely important and such studies are imperative. Snake venom contains several organic and inorganic compounds; phospholipases A2 (PLA2s) are one of the principal toxic components of venom. PLA2s display a wide variety of pharmacological activities, such as neurotoxicity, myotoxicity, cardiotoxicity, anticoagulant, hemorrhagic, and edema-inducing effects. PLA2 inhibition is of pharmacological and therapeutic interests as these enzymes are involved in several inflammatory diseases. This review describes the results of several studies of plant extracts and their isolated active principles, when used against crude snake venoms or their toxic fractions. Isolated inhibitors, such as steroids, terpenoids, and phenolic compounds, are able to inhibit PLA2s from different snake venoms. The design of specific inhibitors of PLA2s might help in the development of new pharmaceutical drugs, more specific antivenom, or even as alternative approaches for treating snakebites.
This datasheet on Schizolobium parahyba covers Identity, Overview, Associated Diseases, Pests or Pathogens, Distribution, Dispersal, Biology & Ecology, Environmental Requirements, Natural Enemies, Impacts, Uses, Management, Genetics and Breeding, Further Information.
Snakebite envenoming is a potentially fatal disease categorized as a neglected public health issue for not receiving the appropriate attention from national and international health authorities. The most affected people by this problem usually live in poor rural communities, where medical resources are often sparse and, in some instances, there is even a scarcity of serum therapy. The administration of the appropriate antivenom is the only specific treatment available, however it has limited efficacy against venom-induced local effects. In this scenario, various plant species are used as local first aid for the treatment of snakebite accidents in Brazil, and some of them can effectively inhibit lethality, neurotoxicity, hemorrhage, and venom enzymes activities. This review compiles a list of plants used in the treatment of snakebites in Brazil, focusing on the native Brazilian species registered in the databases Pubmed, Scielo, Scopus and Google Scholar. All these searches were limited to peer-reviewed journals written in English, with the exception of a few articles written in Portuguese. The most cited native plant species were Casearia sylvestris Sw., Eclipta prostrata (L.) L., Mikania glomerata Spreng., Schizolobium parahyba (Vell.) S.F.Blake and Dipteryx alata Vogel, all used to decrease the severity of toxic signs, inhibit proteolytic and hemorrhagic activities, thus increasing survival time and neutralizing myotoxicity effects. Different active compounds showing important activity against the snake venoms and their toxins include flavonoids, alkaloids and tannins. Although some limitations to the experimental studies with medicinal plants were observed, including lack of comparison with control drugs and unknown active extracts compounds, species with anti-venom characteristics are effective and considered as candidates for the development of adjuvants in the treatment of snake envenomation. Further studies on the chemistry and pharmacology of traditionally used plant species will help to understand the role that snakebite herbal remedies may display in local medical health systems. It might also contribute to the development of alternative or complementary treatments to reduce the number of severe disabilities and deaths.
Snakebites have been declared a neglected health problem that must be considered a national disease of the WHO[world health organisation]. Asian countries like India have high snakebite death rates due to short antidotes and poorly equipped doctors. In today's scenario, local resources like herbs need to be used to prepare cheap antidotes and often available to victims. Snake bites should be viewed as an emergency problem and require additional national guidelines, doctor training, expertise, and human concentration for effective and timely treatment-measures to be taken to ensure the availability and mass production of antidotes. Currently available, antidotes have problems with storage, manufacture, and aspects of the results. Attention should be paid to the natural compound Gedunin with antitoxic effects. To determine Gedunin's therapeutic efficacy well-designed clinical research is required. This article emphasizes and proves the therapeutic effectiveness of the herbal plant active ingredient Gedunin against snakebites.
Snakebites are a serious public health problem. In recent years, several studies have been published that give pharmacological evidence on the benefits of certain species of plants against the effects of snakebites. This review shows an updated list of plants popularly used as antivenoms around the world.
Systemic envenomation by Crotalus durissus terrificus (South American rattlesnake) can cause coagulopathy, rabdomyolysis, acute kidney injury, and peripheral neuromuscular blockade, the latter resulting in flaccid paralysis. Previous studies have shown that plant products such as tannic acid and theaflavin can protect against the neuromuscular blockade caused by C. d. terrificus venom in vitro. In this work, we used mouse-isolated phrenic nerve-diaphragm preparations to examine the ability of caffeic acid, chlorogenic acid, and quercetin to protect against C. d. terrificus venom-induced neuromuscular blockade in vitro. In addition, the ability of tannic acid to protect against the systemic effects of severe envenomation was assessed in rats. Preincubation of venom with caffeic acid (0.5 mg/mL), chlorogenic acid (1 mg/mL), or quercetin (0.5 mg/mL) failed to protect against venom (10 μg/mL)-induced neuromuscular blockade. In rats, venom (6 mg kg−1, i.p.) caused death in ~8 h, which was prevented by preincubation of venom with tannic acid or the administration of antivenom 2 h post-venom, whereas tannic acid given 2 h post-venom prolonged survival (~18.5 h) but did not prevent death. Tannic acid (in preincubation protocols or given 2 h post-venom) had a variable effect on blood creatinine and urea and blood/urine protein levels and prevented venom-induced leukocytosis. Tannic acid attenuated the histological lesions associated with renal damage in a manner similar to antivenom. The protective effect of tannic acid appeared to be mediated by interaction with venom proteins, as assessed by SDS-PAGE. These findings suggest that tannic acid could be a potentially useful ancillary treatment for envenomation by C. d. terrificus.
Medicinal plants are known to possess pharmacologically active compounds which have therapeutic properties and number of plants have been evaluated in disease management including venomous bites. Products of plant extracts are thus gaining importance because of their easy availability, low cost and less side effects and are evaluated for their antidote potential against Indian cobra (Naja naja) venom enzymes. In vitro enzyme inhibitions such as anti-proteolytic, anti-phospholipase, anti-hyaluronidase, anti-acetylcholine esterase and pharmacological studies (anti- hemolytic activity and anti-fibrinogenolytic activity) were evaluated for aqueous ethanolic extracts and organic extracts of Clerodendrum serratum, Azadirachta indica, Aegle marmelos, Aristolochia indica, Citrus limon, Calotropis gigantea, Cryptolepis buchanani and Butea monosperma on N. naja venom. Significant inhibitions were observed by both the extracts. However, it is interesting to note that aqueous extracts significantly inhibited phospholipase activity and organic extracts inhibited fibrinogenolytic activity of venom. The hyaluronidase activity was not observed in the venom. Indirect hemolytic assay revealed the potential of Butea monosperma extracts to completely neutralize the lysis of red blood cells (RBC) induced by cobra venom. The isolation of bioactive principles or a combination of these active principles, from the tested medicinal plants for ex vivo and in vivo anti-venom activity has to be validated by active isolated compound from extract which is responsible for neutralizing toxic snake venom enzymes.
De acuerdo a la Organización Mundial de la Salud más de 1 billón de personas distribuidas en 149 países son afectadas por enfermedades tropicales desatendidas, ocasionando cuantiosos daños económicos, sociales y psicológicos a las personas afectadas, así como un elevado gasto estatal. El envenenamiento por mordedura de serpiente es una de las más desatendidas: se estima que anualmente de los casi 5 millones de mordeduras de serpiente que ocurren a nivel mundial, la mitad genera envenenamientos que ocasionarían entre 94-125 mil muertes, 400 mil amputaciones y otras secuelas severas. Por ello se realizó una búsqueda en Pubmed para identificar publicaciones en los que se hayan usado terapias complementarias o tradicionales o alguno de sus componentes. De los 142 artículos, 18 artículos fueron seleccionados por tratarse de estudios in-vivo para identificar el efecto antiofídico de los compuestos. Los estudios seleccionados se enfocaron en evaluar el efecto antihemorrágico (13/18), anti-edematoso (11/18), anti-necrotizante (5/18) y de reducción de letalidad (4/18). Se estudió el efecto de los compuestos en veneno de Bothrops atrox (6/18), Bothrops jararaca (5/18), Bothrops asper (3/18), Bothrops jararacussu (2/18), Bothrops erythromelas (2/18), Bothrops paulensis (1/18), Crotalus adamanteus (1/18), Crotalus durissus terrificus (1/18), y Lachesis muta (1/18). De las 24 plantas evaluadas, se encontró mayor cantidad de publicaciones sobre el efecto terapéutico de Bellucia dichotoma, Connarus favosus, Plathymenia reticulata, Jatropha gossypiifolia y Renealmia alpinia.
Snakebites are a serious problem in public health due to their high morbimortality. Most of snake venoms produce intense local tissue damage, which could lead to temporary or permanent disability in victims. The available specific treatment is the antivenom serum therapy, whose effectiveness is reduced against these effects. Thus, the search for complementary alternatives for snakebite treatment is relevant. There are several reports of the popular use of medicinal plants against snakebites worldwide. In recent years, many studies have been published giving pharmacological evidence of benefits of several vegetal species against local effects induced by a broad range of snake venoms, including inhibitory potential against hyaluronidase, phospholipase, proteolytic, hemorrhagic, myotoxic, and edematogenic activities. In this context, this review aimed to provide an updated overview of medicinal plants used popularly as antiophidic agents and discuss the main species with pharmacological studies supporting the uses, with emphasis on plants inhibiting local effects of snake envenomation. The present review provides an updated scenario and insights into future research aiming at validation of medicinal plants as antiophidic agents and strengthens the potentiality of ethnopharmacology as a tool for design of potent inhibitors and/or development of herbal medicines against venom toxins, especially local tissue damage.
Detoxification effect of aqueous, methanol and petroleum ether extracts of medicinal plants such as Aristolochia bracteolata, Mucuna pruriens, Prosopis cineraria and Rauvolfia tetraphylla was systematically screened against lethality of crude venom of Naja naja using Swiss albino mice as animal models. We have herein demonstrated that aqueous bark extract of P. cineraria has substantial anti-venom potential vis-a-vis other extracts used in the present study. The aqueous extract at the dose of 14 mg/kg b.w. was able to almost completely neutralize the lethal activity of 3LD 50 (1.12 mg/kg b.w.) of the cobra venom and the extract did not cause any types of adverse side-effects to the animal models. The investigation justifies not only the veraciousness of the extract used by traditional healers of Asian subcontinent as antidotes to snake venoms and also suggests that the aqueous extract should contain specific inhibitors to most principle toxic components of the crude venom. DOI: http://dx.doi.org/10.3329/bjp.v8i4.16684 Bangladesh Journal of Pharmacology Vol.8(4) 2013 395-400
Essential oils are pharmacologically active and unexplored compounds. The inhibitory properties of essential oils from Baccharis dracunculifolia, Conyza bonariensis, Tithonia diversifolia and Ambrosia polystachya were evaluated in the coagulation and fibrinogenolysis induced by snake venoms. The essential oil from Conyza bonariensis extended the clotting time of Lachesis muta from 52.2 to 115.2 seconds and that of Bothrops moojeni from 108.3 to 2340.0 seconds, when pre-incubated with the venoms. The longest clotting times for Bothrops atrox venom were observed after incubation with the essential oils from Conyza bonariensis and Tithonia diversifolia: the times increased from 100.8 to 264.0 and 227.7 seconds, respectively. The prior incubation of the essential oils with plasma and subsequent addition of Lachesis muta venom resulted in a pro-clotting effect. The oils from Ambrosia polystachya and Baccharis dracunculifolia caused 100% of inhibition on the fibrinogenolysis induced by Bothrops moojeni and Lachesis muta venoms (the oils were previously incubated with the venom). The results indicate that the essential oils show promise as adjuvants for the treatment of snakebites.
Envenomations with snakes Bothrops genus can cause dependency at the sting site, which are not reversed even after treatment with snake antivenoms. Incubation of the aqueous extract of Hedychium coronarium (Zingeberaceae) with snake venom Bothrops pauloensis in different concentrations was able to inhibit some enzymatic activities. This work has set a model of regression between concentrations of extract and clotting time (seconds). The adjusted model has captured about 96% of the total variation of clotting time.
Ophidian accidents constitute a serious problem of public health in the tropical countries. In Central and South America, most of the accidents are caused by Bothrops (90.5%), followed by the Crotalus (7.7%), Lachesis (1.4%) and Micrurus (0.4%) genus. The aim of this work was to evaluate clinical-epidemiological aspects of ophidian accidents reported and treated at the Clinical Hospital at Federal University of Uberlândia, in the central region of Brazil. In this study, 641 medical records from January 1999 to December 2013 were analyzed. The results showed that the accidents were more common in the afternoon, from October to April. The major bite occurrence frequency was attributed to the Bothrops (54.76%), followed by Crotalus (30.58%) and Micrurus (1.40%) snakes. Most of the victims were males (80.34%). The main anatomical regions bitten were the lower and upper limbs, 65.67% and 30.58%, respectively. Approximately 80% of the victims were treated in the first 6 hours after the accident. © 2014, Universidade Federal de Uberlandia. All Rights Reserved.
Olax viridis (Olacaceae) and Syzygium guineense (Myrtaceae) are shrubs commonly found in the tropics. They are traditional folkloric medicine for a great number of sicknesses. Olax viridis has a wide range of applications in ethnomedicine which include treatment for ulcers, veneral diseases, ringworm, sleeping sickness, diarrhea, fever. Syzygium guineense has been reported as antidiarrheal agent. Liquid from the bark and roots have been reported to act as a purgative when mixed with water. Both plants have been claimed to have antivenom properties. However, there are no scientific reports on snake venom neutralizing activities of these plants. The plant samples were collected from Olowa in Dekina Local Government Area in Kogi State, Nigeria. The chemicals and reagents used were of analytical grade. Wistar albino rats (male) weighing between 180-200 g were randomly divided into seven groups of three (3). Groups 1-7 received water, normal saline, venom, venom and Olax viridis, venom and Syzygium guineense, Olax viridis and Syzygium guineense respectively. The extracts were administered orally at the dose of 400 mg kg-1 b.w of rats and 1 h later, the venom (0.08 mk kg-1) was administered. Pulse rate, blood glucose, rectal temperature, plasma cholesterol, triacylglycerol, creatine kinase activity and edema were measured. Significant neutralization of the effects of Naja katiensis venom was observed in the groups of rats that received the extracts. Blood glucose, pulse rate, rectal temperature and creatine kinase activity were elevated in the untreated envenomated groups. These results suggest that oral administration of Olax viridis and Syzygium guineense extracts possess antivenom property, thus, providing the rationale for their use in treatment of sake envenomation.
Detoxification effect of aqueous, methanol and petroleum ether extracts of medicinal plants such as Aristolochia bracteolata, Mucuna pruriens, Prosopis cineraria and Rauvolfia tetraphylla was systematically screened against lethality of crude venom of Naja naja using Swiss albino mice as animal models. We have herein demonstrated that aqueous bark extract of P. cineraria has substantial anti-venom potential vis-a-vis other extracts used in the present study. The aqueous extract at the dose of 14 mg/kg b.w. was able to almost completely neutralize the lethal activity of 3LD50 (1.12 mg/kg b.w.) of the cobra venom and the extract did not cause any types of adverse side-effects to the animal models. The investigation justifies not only the veraciousness of the extract used by traditional healers of Asian subcontinent as antidotes to snake venoms and also suggests that the aqueous extract should contain specific inhibitors to most principle toxic components of the crude venom.
Venomous snakebite has been a major cause of mortality and morbidity across the Asian, African and Latin American countries. Lack of medical infrastructure, ineffectiveness of conventional antivenin and malpractice by the local quacks worsen the scenario. The present review deals with the pharmacological investigations performed in different botanicals for antiophidian principles. It also includes a list of certain traditionally used medicinal plants with potential anti snake venom efficacy. The authors have compiled a number of plants active in vitro and/or in vivo against the toxicity of various snake venoms causing an array of biological symptoms. This review also compiles the information regarding the possible use of plant derived natural product based antivenins in order to find cheap and effective alternative source of snake venom antidote especially for the third world tropical countries. From a variety of literature sources the data has been collected mentioning the plants alphabetically and their respective families with notes on plant parts and solvent system used, in vitro and in vivo analyses, activity against the toxicity and biological symptoms related to poisonous snakebite, dose dependence, experimental models, efficacy of the isolated compound(s), ethnobotanical and clinical relevance etc.
The morbidity and mortality associated with snake bite is a serious health concerned especially in developing countries. The rural communities are the worst affected and where it is considered as one of the occupational health hazards especially related to agriculture industry. About 50,000 deaths are recorded per year as a result of snakebite. The scenario may sometimes deteriorate further because of Ineffectiveness and or complications of the anti snake venom as well as untimely interventions or lack of appropriate medications in venomous snakebite cases. The common poisonous snakes found in India are Cobra (Naja naja), Krait (Bangarus Caeruleus), Russell’s viper (Daboia russelli) and saw scaled viper (Echis Carinatus). Over the years many attempts have been made for the development of snake venom antagonists from plants sources. Ethnobotanical data suggests that certain plant species are used traditionally all over the world to treat snakebite cases successfully. Randomness or the use of a variety of species in different families appears to be a feature of traditional snake bite treatments. Present article deals with the Traditional, ethno botanical and pharmacological review of certain plants utilized in the cases of snakebite.
In spite of vast advances in healthcare services, treatment of snakebite still remains a challenge to medical fraternity, because of unresolved complications of severe local tissue damage and consequential physical disabilities. Though anti-venom therapy reduces mortality, is ineffective against local tissue damage. In vitro and in vivo studies demonstrated that several alkaloids, flavonoids, polyphenols, terpenoids, saponins, sterols, glycosides, etc., from herbal medicines effectively neutralized local tissue damage induced by venom toxins/enzymes. This review emphasizes the interplay of venom toxins/enzymes in local toxicity and their neutralization using phytochemicals. Further, approaches using phytochemicals and anti-venoms are reviewed for better management of snakebite.
Read More: http://informahealthcare.com/doi/abs/10.3109/15569543.2013.854255
To elucidate the anti-venom mechanism of persimmon tannin, the interaction between a polymeric persimmon proanthocyanidin fraction (PT40) and phospholipase A2 (PLA2) or bovine serum albumin (BSA) were studied using a competitive binding assay and spectroscopic methods including Fourier transform infrared spectroscopy (FT-IR), circular dichroism (CD), and resonance light scattering (RLS) spectroscopy. The results revealed that PT40 has a higher affinity for PLA2 than for BSA at physiological pH and induced greater conformational changes in PLA2 than in BSA. PT40 covalently bound to PLA2 in a reaction probably involving Lys residues. We propose that the high affinity of PT40 for PLA2 and the covalent modification of PLA2 by PT40 may be responsible for the ability of the tannin to irreversibly inhibit PLA2 catalytic activity, to prevent edema, and to neutralize the lethality of Chinese cobra PLA2in vivo.
The mortality associated with snake bites is a serious public health problem as the estimated death incidence per year is about 1,25,000 globally. In India about 35,000 to 50,000 people reportedly die of snake bite; although, unreported cases may be even more in rural areas. Considering the socio-medical problem due to snake bite, a review is being conducted on snake bite (management aspects), snake venom (nature and its utility), anti-venom and herbal antidote to provide adequate information to researchers for better future prospective.
Envenomation causes an estimated 1.8-2.5 million incidences per year with a mortality level of 100-125,000 persons annually and more than 100,000 individuals suffer from severe complications, which may end in amputation of the attacked limb. The use of plants is a major part of the traditional practitioners' treatment of snakebites.
A database was created for plants used to treat snakebites worldwide. From this database, we selected five countries with a high number of entries and representing different cultures, geography and floristic zones: Brazil, Nicaragua, Nepal, China and South Africa. The datasets were analysed by regression and binominal analysis to see if any family or genus used against snakebites was overrepresented in the respective traditional medicinal systems relative to the abundance in the local flora. The families from the different geographical areas were compared to ascertain whether the same plant families are preferred by different peoples.
Three 'hot' families (Apocynaceae, Lamiaceae and Rubiaceae) were recovered in at least two of the five compared countries in the regression analyses and one 'hot' family (Zingiberaceae) was recovered in two of the compared countries in the binomial analyses. Four out of five floras possess families identified as outliers in both regression and binomial analyses. Eight families were recovered by both the binomial and the regression analysis (40-62% of all highlighted families respectively). At the genus level, only Piper (Piperaceae) was recovered as a 'hot' genus in at least two floras. Seven genera were highlighted by both analyses (25-44% of the highlighted genera).
Cross-cultural comparison of medicinal floras used against snakebites appears to be useful for highlighting candidate families and genera for further studies.
Four compounds (isoquercitrin, myricetin-3-O-glucoside, catechin and gallocatechin) were isolated from lyophilized aqueous extract of Schizolobium parahyba leaves by chromatography on Sephadex LH-20, followed by semipreparative HPLC using a C-18 column, and identified by 1H and 13C NMR. The compounds were then, tested against hemorrhagic and fibrinogenolytic activities of Bothrops crude venoms and isolated metalloproteinases. The inhibitors neutralized the biological and enzymatic activities of Bothrops venoms and toxins isolated from B. jararacussu and B. neuwiedi venoms. The results showed that gallocatechin and myricetin-3-O-glucoside are good inhibitors of hemorrhagic and fibrinogenolytic activities of metalloproteinases, respectively. Gallocatechin also inhibited the myotoxic activity of both B. alternatus venom and BnSP-6 (Lys49 PhospholipaseA2 from B. neuwiedi). Circular dichroism and docking simulation studies were performed in order to investigate the possible interaction between BnSP-6 and gallocatechin. This is the first time these compounds and their anti-ophidian properties are reported for S. parahyba species. Forthcoming studies involving X-ray co-crystallization, will be of great importance for the development of new therapeutic agents for the treatment of ophidian accidents and for the better understanding of the structure/function relationship of venom toxins.
Snake bite, a major socio-medical problem of south east asian countries is still depending on the usage of antisera as the one and only source of treatment, which has its own limitations. In India, mostly in rural areas, health centres are inadequate and the snake bite victims mostly depend on traditional healers and herbal antidotes, as an alternative treatment. The present review has been focussed on the varied folk and traditional herbs and their antisnake venom compounds, which might be a stepping stone in establishing the future therapy against snake bite treatment and management.
Resumo: O presente trabalho teve como objetivo avaliar os processos diretos e seqüenciais nos tempos de 1, 2, 3, 4, 6, 24 e 48 horas das extrações de galactomananas de sementes de bracatinga – GM (Mimosa scabrella), barbatimão – GS (Stryphnodendron adstringens) e guapuruvu – GG (Schizolobium parahybae). A eficiência das extrações foi avaliada através das análises por cromatografia de exclusão estérica de alta performance (HPSEC) acoplada aos detectores de espalhamento de luz laser com multiângulos (MALLS), índice de refração (RI) e viscosimétrico (VIS), (HPSECMALLS/ IR/VIS). As extrações diretas de alactomananas de bracatinga apresentaram relação manose:galactose que variou de 1,1:1 a 1,2:1, para os polissacarídeos de barbatimão a variação foi de 2,0:1 a 2,6:1, enquanto para as extrações de uapuruvu a variação foi de 2,2:1 a 2,6:1. Para as extrações seqüenciais, a variação de galactomananas de bracatinga foi de 1,1:1 a 1,2:1, as galactomananas de barbatimão variaram de 2,0:1 a 2,3:1 e para os polissacarídeos de guapuruvu a variação ocorreu entre 2,5:1 e 4,2:1. As extrações diretas correspondentes aos tempos de 1 hora (GM1d), 6 horas (GS6d) e 2 horas (GG2d) para bracatinga, barbatimão e uapuruvu, respectivamente, foram as frações que apresentaram a melhor eficiência nos processos de extração, considerando rendimento e resultados obtidos por HPSECMALLS/ RI/VIS, tais como massa molecular ponderal média (Mw), índice de polidispersão (Mw/Mn), raio de giro (Rg), taxa de recuperação (%) e viscosidade intrínseca ([]w). Os rendimentos e valores de Mw das extrações escolhidas resultaram em 22,2% e 1,08x106 g/mol, 18,3% e 1,15x106 g/mol e 24,9% e 0,94x106 g/mol respectivamente para GM1d, GS6d e GG2d. Os valores de Mw encontrados corresponderam aos maiores valores de Rg e []w na maioria das extrações analisadas. As distribuições de massas moleculares apresentaram concordância com os resultados de Mw, pois evidenciaram elevadas massas moleculares nos tempos iniciais e degradação principalmente nos tempos de 24 e 48 horas de extração. Os resultados para as extrações diretas de 24 horas apresentaram as variações de 1,5x105 a 7x106 g/mol para bracatinga, 9x104 a 5,5x106 g/mol para barbatimão e 2x104 a 3x106 g/mol para guapuruvu. Os resultados obtidos permitiram verificar que cada fonte vegetal de galactomanana apresenta um tempo de extração direta eficaz relativa ao rendimento, ao comportamento macromolecular em solução e às características físico-químicas. Os resultados também favoreceram a aplicação industrial desses polissacarídeos como potenciais fontes de galactomananas.
The true global incidence of envenomations and their severity remain largely misunderstood, except for a few countries where these accidents are rare or are correctly reported. Nevertheless, this information is essential for drawing up guidelines for dealing with snake-bites, to plan drug supplies, particularly antivenin, and to train medical staff on snake-bite treatments. Since the comprehensive review by Swaroop & Grab in 1954 no global survey has been carried out on snake-bite epidemiology. The present article is an attempt to draw the attention of health authorities to snake envenomations and urges them to prepare therapeutic protocols adapted to their needs.
At the Hospital of Clinics of the Faculty of Medicine of Ribeirão Preto/USP during the years 1980-1989 21 children have been attended after rattlesnake bite: 16 severe and 5 with moderate envenomation. Four (20%) developed acute tubular necrosis 2 necessitating dialysis. One patient died 13 days after the bite and grave complications including digestive hemorrhage and acute respiratory insuficiency. All patients preserved clinical laboratory and epidemiological characteristics of Crotalus durissus terrificus envenomation. We also comment on the correct management of such patients specially related to antivenin dosage and the prevention ofacute tubular the most serious complication of such an accident.
The crude aqueous extract from the leaves of Casearia sylvestris, a plant found in Brazilian open pastures, was assayed for its ability to inhibit phospholipase A2 (PLA2) activity and some biological activities of bee and several snake venoms, and of a number of isolated PLA2s. The extract induced partial inhibition of the PLA2 activity of venoms containing class I, II and III PLA2s. When tested against the purified toxins, it showed the highest efficacy against class II PLA2s from viperid venoms, being relatively ineffective against the class I PLA2 pseudexin. In addition, C. sylvestris extract significantly inhibited the myotoxic activity of four Bothrops crude venoms and nine purified myotoxic PLA2s, including Lys-49 and Asp-49 variants. The extract was able to inhibit the anticoagulant activity of several isolated PLA2s, with the exception of pseudexin. Moreover, it partially reduced the edema-inducing activity of B. moojeni and B. jararacussu venoms, as well as of myotoxins MjTX-II and BthTX-I. The extract also prolonged the survival time of mice injected with lethal doses of several snake venoms and neutralized the lethal effect induced by several purified PLA2 myotoxins. It is concluded that C. sylvestris constitutes a rich source of PLA2 inhibitors.
The article surveys the substances identified in plants reputed to neutralize the effects of snake venoms. Protective activity of many of them against the lethal action of the venom of the jararaca (Bothrops jararaca) snake was confirmed by biological assays. It was shown that all belong to chemical classes capable of interacting with macromolecular targets--receptors and enzymes. In a few cases it has been shown that exogenous natural micromolecules can mimic the biological activity of endogenous macromolecules. From the evidence presented, it can be inferred that micromolecules which neutralize the action of snake venoms mechanistically replace endogenous antitoxic serum proteins with venom neutralizing capacity such as produced by some animals.
Aqueous extract from Casearia sylvestris leaves, a typical plant from Brazilian open pastures, was able to neutralize the hemorrhagic activity caused by Bothrops asper, Bothrops jararacussu, Bothrops moojeni, Bothrops neuwiedi and Bothrops pirajai venoms. It also neutralized two hemorrhagic metalloproteinases from Bothrops asper venom. Proteolytic activity on casein induced by bothropic venoms and by isolated proteases, including Bn2 metalloproteinase from B. neuwiedi venom, was also inhibited by the C. sylvestris extract in different levels. The alpha-fibrinogen chain was partially protected against degradation caused by B. jararacussu venom, when this venom was incubated with C. sylvestris extract. We also observed that this extract partially increased the time of plasma coagulation caused by B. jararacussu, B. moojeni and B. neuwiedi venoms. C. sylvestris extract did not induce proteolysis in any substrate assayed.
The peptidomimetic hydroxamate metalloproteinase inhibitor batimastat (BB-94) was assessed for its ability to neutralize the systemic effects (lethality, hemorrhage and coagulopathy) induced by the venom of Bothrops asper, the most important snake from a medical standpoint in Central America. Batimastat inhibited lethality when a venom challenge dose of two LD(50)s was used by intraperitoneal and intravenous routes, with ED(50)s of 250 and 22 microM, respectively. With a challenge dose of three LD(50)s, lethality was not abrogated, but a conspicuous and dose-dependent delay in the time of death was observed in mice injected with mixtures of venom plus batimastat. Upon incubation with 500 microM batimastat, venom LD(50) increased 2.86-fold (intraperitoneal route) and 2.37-fold (intravenous route), when compared with LD(50) of venom alone. Batimastat also inhibited the hemorrhagic effect induced by venom in the lungs after intravenous injection. Moreover, batimastat exerted a significant inhibition of in vitro coagulant and in vivo defibrinogenating effects of venom, evidencing that metalloproteinases play a key role in the coagulopathy characteristic of B. asper envenomation. The remaining uninhibited coagulant effect is due to serine proteinases, i.e. thrombin-like enzymes, since this effect was completely abrogated by the combination of batimastat and PMSF. Our results stress the view that metalloproteinases play a relevant role in the systemic pathophysiology of B. asper envenomation and that metalloproteinase inhibitors may become a therapeutic alternative in this pathology.
A protein determination method which involves the binding of Coomassie Brilliant Blue G-250 to protein is described. The binding of the dye to protein causes a shift in the absorption maximum of the dye from 465 to 595 nm, and it is the increase in absorption at 595 nm which is monitored. This assay is very reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 1 hr. There is little or no interference from cations such as sodium or potassium nor from carbohydrates such as sucrose. A small amount of color is developed in the presence of strongly alkaline buffering agents, but the assay may be run accurately by the use of proper buffer controls. The only components found to give excessive interfering color in the assay are relatively large amounts of detergents such as sodium dodecyl sulfate, Triton X-100, and commercial glassware detergents. Interference by small amounts of detergent may be eliminated by the use of proper controls.
Medicinal plants constitute a very rich source of natural inhibitors of animal toxins, and may be used: 1) to study the mechanism of action of toxins and inhibitors; 2) to treat ophidian envenomation as a supplementary and/or alternative therapy; and 3) as models to design new drugs of interest in clinical medicine. Several Brazilian plants have been utilized in folk medicine as antiophidians. However, only a few species have been scientifically investigated and still fewer have had their active components isolated and characterized both structurally and functionally. This article presents a review of Brazilian species showing neutralizing properties against snake venoms which have been assayed in the research laboratory and characterized ethnopharmacologically in terms of: 1) the part of the plant used as antidote; 2) the respective genus and family; and 3) the main pharmacological properties related to inhibition of toxic and enzymatic activities of snake venoms and isolated toxins.
Phospholipases A2 constitute the major components from Bothrops snake venoms and have been extensively investigated not only because they are relatively very abundant in these venoms but mainly because they display a range of many relevant biological effects, including: myotoxic, cytotoxic, edemainducing, artificial membrane disrupting, anticoagulant, neuromuscular, platelet aggregation inhibiting, hypotensive, bactericidal, anti-HIV, anti-tumoural, anti-malarial and anti-parasitic. The primary structures of several PLA2s have been elucidated through direct amino acid sequencing or, inderectly, through the corresponding nucleotide sequencing. Two main subgroups were thus described: (i) Asp49 PLA2s, showing low (basic, highly myotoxic) to relatively high (acidic, less or non myotoxic) Ca++-dependent hydrolytic activity upon artificial substrates; (ii) Lys49 PLA2s (basic, highly myotoxic), showing no detectable hydrolytic activity on artificial substrates. Several crystal structures of Lys49 PLA2s from genus Bothrops have already been solved, revealing very similar fold patterns. Lack of catalytic activity of myotoxic Lys49- PLA2s, first related solely with the fact that Lys49 occupies the position of the calcium ion in the catalyticly active site of Asp49 PLA2s, is now also attributed to Lys122 which interacts with the carbonyl of Cys29 hyperpolarising the peptide bond between Cys29 and Gly30 and trapping the fatty acid product in the active site, thus interrupting the catalytic cycle. This hypothesis, supported for three recent structures, is also discussed here. All Asp49 myotoxins showed to be pharmacologically more potent when compared with the Lys49 variants, but phospholipid hydrolysis is not an indispensable condition for the myotoxic, cytotoxic, bactericidal, anti-HIV, anti-parasitic, liposome disrupting or edema-inducing activities. Recent studies on site directed mutagenesis of the recombinant Lys49 myotoxin from Bothrops jararacussu revealed the participation of important amino acid residues in the membrane damaging and myotoxic activities.
Crotoxin B, the basic Asp49-PLA2 subunit from crotoxin, the main component of Crotalus durissus terrificus venom, displays myotoxic, edema-inducing, bactericidal (upon Escherichia coli), liposomal-disrupting and anticoagulant activities. Chemical modifications of His (with 4-bromophenacyl bromide, BPB), Tyr (with 2-nitrobenzenesulphonyl fluoride, NBSF), Trp (with o-nitrophenylsulphenyl chloride, NPSC) and Lys (with acetic anhydride) residues of this protein, in addition to cleavage with cyanogen bromide (CNBr) and inhibition with ethylenediaminetetraacetic acid (EDTA), were carried out in order to study their effects on enzymatic and pharmacological activities. Lethality was reduced after modification of His or Lys residues, as well as after cleavage with CNBr, while enzymatic activity was completely abolished after modification of His or incubation with EDTA. Modification of Lys or Tyr, or cleavage with CNBr, partially reduced enzymatic activity. Anticoagulant activity was modified similarly to enzymatic activity, evidencing the dependency of this pharmacological effect on catalytic activity. Myotoxicity was reduced after modification of His or Lys, as well as after cleavage with CNBr, whereas EDTA reduced this effect to a lesser extent. Bactericidal effect was significantly reduced only after modification of Lys and after cleavage with CNBr. Edema-inducing activity was partially inhibited after treatment with EDTA and strongly reduced after acetylation of Lys residues and cleavage with CNBr, being only partially reduced after His alkylation. On the other hand, liposome disrupting activity was only partially reduced after modification of His and Tyr or after cleavage with CNBr. Modification of Trp residue partially reduced lethality and myotoxicity but did not affect enzymatic or anticoagulant activities. These data indicate that enzymatic activity is relevant for some pharmacological effects induced by crotoxin B (mainly lethal, myotoxic and anticoagulant activities), and also evidence that this subunit of crotoxin displays regions different from the active catalytic site which are involved in some of the toxic and pharmacological effects induced by this phospholipase A2.
1.1. Venoms of B. asper, B. atrox, B. marajoensis and B. moojeni collected in different regions were analyzed by their enzymatic activity and polyacrylamide gel electrophoreses (acidic, basic and SDS).2.2. These species can be recognized and distinguished by their characteristic protein electrophoretic patterns.3.3. The venoms of B. atrox from different localities, although having similar protein electrophoretic distribution, display distinct proteinase patterns, indicating the presence of possible subspecies.4.4. By the composition of the venom, the snake B. moojeni must be classified as a species distinct from B. atrox.5.5. The B. asper venoms, from Atlantic and Pacific coasts, have not only different enzymatic activities but also different electrophoretic patterns. The classification of this species must be revised.
A protein determination method which involves the binding of Coomassie Brilliant Blue G-250 to protein is described. The binding of the dye to protein causes a shift in the absorption maximum of the dye from 465 to 595 nm, and it is the increase in absorption at 595 nm which is monitored. This assay is very reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 1 hr. There is little or no interference from cations such as sodium or potassium nor from carbohydrates such as sucrose. A small amount of color is developed in the presence of strongly alkaline buffering agents, but the assay may be run accurately by the use of proper buffer controls. The only components found to give excessive interfering color in the assay are relatively large amounts of detergents such as sodium dodecyl sulfate, Triton X-100, and commercial glassware detergents. Interference by small amounts of detergent may be eliminated by the use of proper controls.
Many plants are recommended in traditional medicine as active against various effects of snakebite. Few attempts have been made to investigate the veracity of these assertions in controlled experiments. Several workers, mainly Oriental, have investigated the reputation of such plants by performing in vitro and in vivo experiments in order to demonstrate whether there was any protective effect, using drugs or mixtures of drugs prepared using traditional formulae. In some studies, these extracts were administered to mice before or after treatment with different elapid or crotalid venoms. Other papers deal with selected compounds isolated from Schumanniophyton magnificum, Eclipta prostrata or Aristolochia shimadai, and their capacity to inhibit phospholipase A2 or other enzymes (e.g. ATPase) or for physiological and biochemical properties (such as effects on uterine tone or the protection of mitochondrial membranes). Japanese workers have described the antihaemorrhagic effect of persimmon tannin from Diospyros kaki. Atropine has been attributed a life-prolonging effect after black mamba venom treatment. Prolonged survival was also observed after pretreatment with extracts of Diodia scandens and Andrographis paniculata. Some authors have found little or no beneficial effects. The papers collected so far show that there are no systematic investigations in this field.
At the Hospital of Clinics of the Faculty of Medicine of Ribeirão Preto/USP during the years 1980-1989 21 children have been attended after rattlesnake bite: 16 severe and 5 with moderate envenomation. Four (20%) developed acute tubular necrosis 2 necessitating dialysis. One patient died 13 days after the bite and grave complications including digestive hemorrhage and acute respiratory insufficiency. All patients preserved clinical laboratory and epidemiological characteristics of Crotalus durissus terrificus envenomation. We also comment on the correct management of such patients specially related to antivenin dosage and the prevention of acute tubular the most serious complication of such an accident.
Changes in the haemostatic mechanism caused by venoms of Bothrops, Crotalus and Lachesis snakes from Central and South America in human accidents are reviewed. Changes in the blood coagulation mechanism could be found depending on the action of the venom on clotting factors.
A review has been presented of the biochemistry and pharmacology of a class of natural products, the flavonoids. These substances which are widely distributed in the plant kingdom and present in considerable quantities in common food products, spices and beverages have in a concentrated form (Propolis) been used since ancient times by physicians and laymen to treat a great variety of human diseases but they have yet to pass the tests of modern, controlled, clinical experimentation. An attempt has been made to present the fundamental evidence from the basic biological sciences which is required to stimulate the interest of the clinicians in this new field. The few existing reports on the careful pharmacodynamic, pharmacokinetic and clinical studies which have been made have been summarized to provide a basis for a full-scale investigation of the therapeutic potential of flavonoids.
Snake bite is an important medical problem in some areas of Papua New Guinea and appears to be most common in the Central Province and National Capital District. The overall incidence for Central Province is 215.5 per 100,000 population, but Kairuku subprovince has an incidence of 526 per 100,000, which is amongst the highest in the world. The clinical pattern of envenoming also varies within the Province, suggesting that different species of snake may be responsible for bites in different areas. Most envenomed patients are bitten during daylight on the lower limb and are rarely able to describe the snake. The mortality rate in Central Province is 7.9 per 100,000; most patients die from ventilatory failure due to severe neurotoxicity. Mortality might be reduced by increased use of compression bandaging as a first aid measure, earlier treatment with antivenom and earlier referral to hospital.
(1) Venom pools from Bothrops neuwiedi (Bn) and from two subspecies, namely Bothrops neuwiedi pauloensis (Bnp) and Bothrops neuwiedi urutu (Bnu), collected in the States of São Paulo (SP) and Minas Gerais (MG), Brazil, were electrophoretically examined. Basic toxins with different isoelectric points were identified in the venom collected in São Paulo (BnSP). These toxins were absent in the corresponding pools from Minas Gerais (BnMG, BnpMG and BnuMG). (2) BnSP, but not BnMG, BnpMG or BnuMG, showed two myotoxins (pI approximately equal to 8.6 and 8.8, respectively) which were isolated by ion-exchange chromatography on CM-Sepharose. (3) From BnMG, three myotoxic isoforms (pI approximately equal to 8.2 and M(r) = 13,600) were isolated by chromatography on CM-Sepharose followed by reversed-phase high-performance liquid chromatography. (4) The chemical and biological characterization of these toxins showed a high similarity with the Lys-49 myotoxins from other bothropic venoms. (5) Doses up to 5 LD50 (i.p.) of p-bromophenacyl bromide alkylated BnSP-7 caused a total loss of lethality in 18-22-g mice, thus indicating that the LD50 was increased by greater than 5-fold. At this dose myotoxicity was also not detectable, but the edematogenic activity on the rat paw apparently did not change.
A fibrino(geno)lytic nonhemorrhagic metalloprotease (neuwiedase) was purified from Bothrops neuwiedi snake venom by a single chromatographic step procedure on a CM-Sepharose column. Neuwiedase represented 4.5% (w/w) of the crude desiccated venom, with an approximate Mr of 20,000 and pI 5.9. As regards the amino acid composition, neuwiedase showed similarities with other metalloproteases, with high proportions of Asx, Glx, Leu, and Ser. Atomic absorption spectroscopy showed that one mole of Zn2+ and one mole of Ca2+ were present per mole of protein. The cDNA encoding neuwiedase was isolated by RT-PCR from venom gland RNA, using oligonucleotides based on the partially determined amino-acid sequences of this metalloprotease. The full sequence contained approximately 594 bp, which codified the 198 amino acid residues with an estimated molecular weight of 22,375. Comparison of the nucleotide and amino acid sequences of neuwiedase with those of other snake venom metalloproteases showed a high level of sequential similarity. Neuwiedase has two highly conserved characteristics sequences H142E143XXH146XXG149XXH152 and C164I165M166. The three-dimensional structure of neuwiedase was modeled based on the crystal structure of Crotalus adamanteus Adamalysin II. This model revealed that the zinc binding site region showed a high structural similarity with other metalloproteases. The proteolyitc specificity, using the Bbeta-chain of oxidized insulin as substrate, was shown to be directed to the Ala14-Leu15 and Tyr16-Leu17 peptide bonds which were preferentially hydrolyzed. Neuwiedase is a Aalpha,Bbeta fibrinogenase. Its activity upon the Aalpha chain of fibrinogen was detected within 15 min of incubation. The optimal temperature and pH for the degradation of both Aalpha and Bbeta chains were 37 degrees C and 7.4-8.0, respectively. This activity was inhibited by EDTA and 1,10-phenantroline. Neuwiedase also showed proteolytic activity upon fibrin and some components of the extracellular matrix. However, it did not show TAME esterase activity and was not able to inhibit platelet aggregation.
The biochemical characteristics of hemorrhagic metalloproteinases isolated from snake venoms are reviewed, together with their role in the pathogenesis of the local tissue damage characteristic of crotaline and viperine snake envenomations. Venom metalloproteinases differ in their domain structure. Some enzymes comprise only the metalloproteinase domain, others have disintegrin-like and high cysteine domains and others present, besides these domains, an additional lectin-like subunit. All of them are zinc-dependent enzymes with highly similar zinc binding environments. Some metalloproteinases induce hemorrhage by directly affecting mostly capillary blood vessels. It is suggested that hemorrhagic enzymes cleave, in a highly selective fashion, key peptide bonds of basement membrane components, thereby affecting the interaction between basement membrane and endothelial cells. As a consequence, these cells undergo a series of morphological and functional alterations in vivo, probably associated with biophysical hemodynamic factors such as tangential fluid shear stress. Eventually, gaps are formed in endothelial cells through which extravasation occurs. In addition to hemorrhage, venom metalloproteinases induce skeletal muscle damage, myonecrosis, which seems to be secondary to the ischemia that ensues in muscle tissue as a consequence of bleeding and reduced perfusion. Microvessel disruption by metalloproteinases also impairs skeletal muscle regeneration, being therefore responsible of fibrosis and permanent tissue loss after snakebites. Moreover, venom metalloproteinases participate in the degradation of extracellular matrix components and play a relevant role in the prominent local inflammatory response that characterizes snakebite envenomations, since they induce edema, activate endogenous matrix metalloproteinases (MMPs) and are capable of releasing TNF-alpha from its membrane-bound precursor. Owing to their protagonic role in the pathogenesis of local tissue damage, snake venom metalloproteinases constitute relevant targets for natural and synthetic inhibitors which may complement antivenoms in the neutralization of these effects.
The pathological alterations induced by neuwiedase, a 22 kDa class P-I metalloproteinase from the venom of the South American pit viper Bothrops neuwiedi, were studied in mice. Neuwiedase was devoid of hemorrhagic activity when tested in the skin up to a dose of 200 microgram, and also after intramuscular injection in the gastrocnemius. However, it induced bleeding when applied onto the mouse cremaster muscle in intravital microscopy experiments, and caused pulmonary hemorrhage when injected intravenously at doses higher than 5 microgram/g. Median lethal dose (LD(50)) by the intravenous route was 5 microgram/g, whereas LD(50) of crude venom was 0.47 microgram/g. After intramuscular injection, neuwiedase induced a mild myotoxic effect, evidenced histologically and by the increment in plasma creatine kinase activity, but it was devoid of hemorrhagic and thrombotic effects. In contrast, crude B. neuwiedi venom induced prominent hemorrhage and myonecrosis in gastrocnemius muscle. Both venom and neuwiedase induced an inflammatory reaction in muscle tissue characterized by abundant polymorphonuclear leukocytes. Moreover, a conspicuous edema developed in the foot pad after subcutaneous injection of neuwiedase. Anti-neuwiedase antibodies produced in rabbits were effective in the neutralization of hemorrhagic activity of crude venom, evidencing immunological cross-reactivity between neuwiedase and other hemorrhagic metalloproteinases present in the venom, and suggesting that metalloproteinases devoid of, or having low, hemorrhagic activity could be good immunogens to generate antibodies effective against high molecular mass metalloproteinasas having potent hemorrhagic activity. It is concluded that neuwiedase, despite its lack of hemorrhagic effect when injected in the gastrocnemius muscle, contributes to local tissue damage by inducing edema, inflammatory infiltrate and mild myotoxicity, and by degrading extracellular matrix components. In addition, large doses of neuwiedase may contribute to pulmonary bleeding
Coagulopathy is one of the major complications following envenomations by crotalid and viperid snakes. The present study was undertaken to examine the effect of a hemorrhagic metalloproteinase in Bothrops jararaca venom, jararafibrase I (JF I), on the development of coagulopathy using rat snakebite model. Coagulation parameters were monitored after subcutaneous injection of B. jararaca crude venom, JF I-neutralized venom and purified JF I in rats. Crude venom induced unclottable blood and fibrinogen consumption, while JF I-neutralized venom and purified JF I did not induce coagulopathy. Plasma venom antigen level of rats given JF I-neutralized venom was lower than that of rats given crude venom. We conclude that venom hemorrhagic metalloproteinases play an important role in the development of coagulopathy through rapid spreading of venom coagulation components from the injected area into systemic circulation.
As a tribute to Revista de Biología Tropical in its 50th anniversary, this review describes some of the main research efforts carried out in the study of the chemical composition and the mechanism of action of toxins present in the venoms of snakes distributed in Latin America. Venom proteins involved in neurotoxicity, coagulopathies, hemorrhage and muscle necrosis are discussed, together with a description of the inflammatory reactions elicited by these venoms and toxins. In addition, the search for inhibitory substances present in plants and animals that may be utilized in the neutralization of venoms is analyzed. Some of the clinical studies performed on snakebite envenomations in Latin America are also reviewed, together with the development of technologies aimed at improving the quality of antivenoms produced in the region. Toxinology has become a fruitful and stimulating research field in Latin America which has contributed to a better understanding of snake venoms as well as to an improved management of snake bitten patients.
The aqueous extract from the leaves of Casearia mariquitensis (C. m.), a plant found in Brazilian open pastures, was assayed for its ability to inhibit some hematological and hemostatic effects induced by neuwiedase, a 22 kDa class P-I metalloproteinase from the venom of the South American pit viper Bothrops neuwiedi pauloensis. The aqueous extract from C. m. was able to neutralize the hematological alterations induced by the crude venom (C.V.) upon erythrocytes when the venom was incubated at a ratio of 1:10 (w/w, venom/extract), but it did not neutralize the platelet decreasing ability of C.V. The plasma fibrinogen concentration decreased approximately 36% and 83% when 0.6 LD(50) of the C.V. or neuwiedase, respectively, were injected by i.p. route in mice, and the aqueous extract from C. m. was able to inhibit this effect. The Bbeta fibrinogen chain was protected against degradation caused by crude venom and neuwiedase when the venom or toxin were incubated with C. m. extract. We also observed that this extract exerted a very slight effect on the clotting time, prolonging it only to a little extent. The pulmonary hemorrhage induced by neuwiedase when injected intravenously with 0.6 LD(50) was completely inhibited when this toxin was incubated with the extract at a ratio of 1:10 (w/w, toxin/extract). It is concluded that C. m. displays components able to inhibit some hematological and systemic alterations induced by C.V.
Phospholipases A(2) (PLA(2)) are multifunctional proteins which exhibit varied biological activities correlated to the structural diversities of the sub-classes. The crude aqueous extract from subterranean system of Mandevilla velutina, a plant found in Brazilian savanna, was assayed for its ability to inhibit biological activities of several snake venoms and isolated PLA(2)s. The extract induced total inhibition of the phospholipase activity of Crotalus durissus terrificus venom and only partial inhibition of Bothrops venoms. When assayed against purified toxins, the highest efficacy was detected against CB and crotoxin, while almost ineffective against PLA(2)s from the genus Bothrops. Although M. velutina crude extract significantly inhibited the myotoxic activity of C. d. terrificus venom and CB, it produced only partial inhibition of either Bothrops jararacussu venom or its main myotoxins BthTX-I (basic Lys49), BthTX-II (basic Asp49) and BthA-I-PLA(2) (acidic Asp49). The extract exhibited also full inhibition of hemorrhage caused by Bothrops alternatus, Bothrops moojeni and Bothrops pirajai snake venoms, but partial inhibition (90%) of that induced by B. jararacussu venom. The extract was ineffective to inhibit the fibrinogenolytic activity of B. moojeni, B. alternatus and B. pirajai crude venoms, while their caseinolytic activity was only partially inhibited. No inhibition of the anticoagulant activity, although partial reduction of the edema-inducing activity of C. d. terrificus and B. alternatus crude venoms, CB, PrTX-I, BthTX-I and crotoxin was observed. Besides extending survival of mice injected with lethal doses of C. d. terrificus and B. jararacussu venoms, M. velutina extract decreased to 50% the lethality of mice. Extracts of 18 month old micropropagated plants were able to partially neutralize the effect of the crude venoms and toxins.
The aqueous extract from aerial parts of Bauhinia forficata was able to neutralize the clotting activity induced by Bothrops and Crotalus crude venoms. The clotting time, upon human plasma, induced by B. moojeni venom was significantly prolonged. Clotting and fibrinogenolytic activities induced by isolated thrombin-like enzyme from Bothrops jararacussu were totally inhibited after incubation at different ratios. The extract was not able to neutralize the hemorrhagic activity induced by an Bothrops venoms, but it efficiently inhibited the edema induced by Crotalus durissus terrificus venom and isolated PLA2s. In addition, it did not inhibited the phospholipase A2 activity of Bothrops snake venoms. Interaction studies between Bauhinia forficata extract and snake venoms, when analyzed by SDS-PAGE, did not reveal any apparent degradation of the venom proteins. This extract is a promising source of natural inhibitors of serine-proteases involved in blood clotting disturbances induced by snake venoms.
The use of plants as medicine has been referred to since ancient peoples, perhaps as early as Neanderthal man. Plants are a source of many biologically active products and nowadays they are of great interest to the pharmaceutical industry. The study of how people of different culture use plants in particular ways has led to the discovery of important new medicines. In this work, we verify the possible activity of Musa paradisiaca L. (Musaceae) against the toxicity of snake venoms. Musa paradisiaca, an important source of food in the world, has also been reported to be popularly used as an anti-venom. Interaction of Musa paradisiaca extract (MsE) with snake venom proteins has been examined in this study. Phospholipase A2 (PLA2), myotoxic and hemorrhagic activities, including lethality in mice, induced by crotalidae venoms were significantly inhibited when different amounts of MsE were mixed with these venoms before assays. On the other hand, mice that received MsE and venoms without previous mixture or by separated routes were not protected against venom toxicity. Partial chemical characterization of MsE showed the presence of polyphenols and tannins and they are known to non-specifically inactivate proteins. We suggest that these compounds can be responsible for the in vitro inhibition of the toxic effects of snake venoms. In conclusion, according to our results, using mice as experimental model, MsE does not show protection against the toxic effects of snake venoms in vivo, but if was very effective when the experiments were done in vitro.
Zinc-dependent metalloproteinases are responsible for the hemorrhagic activity characteristic of viperid snake venoms. Snake venom metalloproteinases (SVMPs) are classified in various groups (P-I-IV), according to their domain composition. P-III SVMPs, comprising metalloproteinase, disintegrin-like and cysteine-rich domains, exert more potent hemorrhagic activity than P-I SVMPs, which present only the metalloproteinase domain. SVMPs degrade various components of the basement membrane and are also able to hydrolyze endothelial cell membrane proteins, such as integrins and cadherins, involved in cell-matrix and cell-cell adhesion. In addition, disintegrin-like and cysteine-rich domains interact with endothelial cell integrins, interfering with their adhesion to extracellular matrix. Hemorrhage induced by SVMPs is an extremely rapid event in vivo, with capillary endothelial cells showing drastic structural alterations within few minutes. In contrast, observations in cell culture conditions do not evidence such rapid endothelial cell damage. Instead, the main effect is detachment and rounding of these cells; it is only after several hours of incubation that cells show evidence of apoptotic damage. This apparent discrepancy between in vivo and in vitro observations can be explained if biophysical forces operating on microvessels in vivo are taken into consideration. It is proposed that SVMP-induced hemorrhage occurs in vivo by a 'two-step' mechanism. Initially, SVMPs degrade basement membrane and adhesion proteins, thus weakening the capillary wall and perturbing the interactions between endothelial cells and the basement membrane. Then, transmural pressure acting on the weakened capillary wall causes distention. As a consequence, endothelial cells become very thin, until the integrity of the capillary wall is lost at some points, where extravasation occurs. In addition, endothelial cells become more susceptible to blood flow-dependent shear stress, which further contributes to capillary wall disruption.
Many plants are used in traditional medicine as active agents against various effects induced by snakebite. The methanolic extract from Cordia verbenacea (Cv) significantly inhibited paw edema induced by Bothrops jararacussu snake venom and by its main basic phospholipase A2 homologs, namely bothropstoxins I and II (BthTXs). The active component was isolated by chromatography on Sephadex LH-20 and by RP-HPLC on a C18 column and identified as rosmarinic acid (Cv-RA). Rosmarinic acid is an ester of caffeic acid and 3,4-dihydroxyphenyllactic acid [2-O-cafeoil-3-(3,4-di-hydroxy-phenyl)-R-lactic acid]. This is the first report of RA in the species C. verbenacea ('baleeira', 'whaler') and of its anti-inflammatory and antimyotoxic properties against snake venoms and isolated toxins. RA inhibited the edema and myotoxic activity induced by the basic PLA2s BthTX-I and BthTX-II. It was, however, less efficient to inhibit the PLA2 activity of BthTX-II and, still less, the PLA2 and edema-inducing activities of the acidic isoform BthA-I-PLA2 from the same venom, showing therefore a higher inhibitory activity upon basic PLA2s. RA also inhibited most of the myotoxic and partially the edema-inducing effects of both basic PLA2s, thus reinforcing the idea of dissociation between the catalytic and pharmacological domains. The pure compound potentiated the ability of the commercial equine polyvalent antivenom in neutralizing lethal and myotoxic effects of the crude venom and of isolated PLA2s in experimental models. CD data presented here suggest that, after binding, no significant conformation changes occur either in the Cv-RA or in the target PLA2. A possible model for the interaction of rosmarinic acid with Lys49-PLA2 BthTX-I is proposed.
Several Brazilian plants have been utilized in folk medicine as active agents against various effects induced by snake venoms. The inhabitants of the Amazon region use, among others, the macerated bark of a plant popularly named "Pracaxi" (Pentaclethra macroloba Willd) to combat these effects. We report now the antihemorrhagic properties against snake venoms of the aqueous extract of Pentaclethra macroloba (EPema). EPema exhibited full inhibition of hemorrhagic and nucleolytic activities induced by several snake venoms. Additionally, partial inhibition of myotoxic, lethal, phospholipase and edema activities of snake venoms and its isolated PLA(2)s by EPema is reported. In vivo tests showed that EPema is able to totally inhibit a Bothrops jararacussu metalloprotease (BjussuMP-I) induced hemorrhage, suggesting interaction of the extract compounds with this high molecular weight protein. The extract did induce neither hemorrhage nor death in mice when administered alone by i.m. route. When administered separately by i.m. route, the extract did not induce death in mice at 12.5--300 mg/kg doses. Other assays demonstrated that EPema was unable to inhibit fibrinogenolytic and coagulant activities of Bothrops atrox venom. Although the mechanism of action of EPema is still unknown, the finding that no visible change was detected in the electrophoretic pattern of snake venom after incubation with the extract excludes proteolytic degradation as a potential mechanism. The search for new inhibitors of venom metalloproteases and DNAases are a relevant task. Investigation of snake venom inhibitors can provide useful tools for the elucidation of the action mechanisms of purified toxins. Furthermore, these inhibitors can be used as molecular models for development of new therapeutical agents in the treatment of ophidian accidents.
Aqueous extracts, prepared from dried or fresh roots, stems or leaves of Mikania glomerata, a plant found in Mata Atlântica in Southeastern Brazil, were able to efficiently neutralize different toxic, pharmacological, and enzymatic effects induced by venoms from Bothrops and Crotalus snakes. Phospholipase A(2) activity and the edema induced by Crotalus durissus terrificus venom were inhibited around 100 and approximately 40%, respectively, although this inhibition was only partial for Bothrops venoms. The hemorrhagic activity of Bothrops venoms (Bothrops altenatus, Bothrops moojeni, Bothrops neuwiedi, and Bothrops jararacussu) was significantly inhibited by this vegetal species, while the clotting activity of Crotalus durissus terrificus, Bothrops jararacussu, and Bothrops neuwiedi venoms was totally inhibited. Although, the mechanism of action of Mikania glomerata extract is still unknown, the finding that no visible change was detected in the electrophoretic pattern of snake venom after incubation with the extract excludes proteolytic degradation as a potential mechanism. Since the extract of Mikania glomerata significantly inhibited the studied snake venoms, it may be used as an alternative treatment to serumtherapy and, in addition, as a rich source of potential inhibitors of PLA(2)s, metalloproteases and serineproteases, enzymes involved in several physiopathological human and animal diseases.
Envenomations due to snake bites are commonly treated by parenteral administration of horse or sheep-derived polyclonal antivenoms aimed at the neutralization of toxins. However, despite the widespread success of this therapy, it is still important to search for different venom inhibitors, either synthetic or natural, that could complement or substitute for the action of antivenoms. Several plants have been utilized in folk medicine as antiophidian. However, only a few species have been scientifically investigated and still less had their active components isolated and characterized both structurally and functionally. This article presents a review of plants showing neutralizing properties against snake venoms which were assayed in research laboratories, correlating them with ethnopharmacological studies, as (i) the part of the plant used as antidote, (ii) its respective genus and family and (iii) inhibition of the main pharmacological, toxic and enzymatic activities of snake venoms and isolated toxins. Protective activity of many of these plants against the lethal action of snake venoms has been confirmed by biological assays. Compounds in all of them belong to chemical classes capable of interacting with macromolecular targets (enzymes or receptors). Popular culture can often help to guide scientific studies. In addition, biotechnological application of these inhibitors, as helpful alternative or supplemental treatments to serum therapy, and also as important models for synthesis of new drugs of medical interest, needs to be better oriented and scientifically explored.
In Indian traditional medicine, various plants have been used widely as a remedy for treating snake bites. The aim of this study was to evaluate the effect of Tamarindus indica seed extract on the pharmacological as well as the enzymatic effects induced by V. russelli venom. Tamarind seed extract inhibited the PLA(2), protease, hyaluronidase, l-amino acid oxidase and 5'-nucleotidase enzyme activities of venom in a dose-dependent manner. These are the major hydrolytic enzymes responsible for the early effects of envenomation, such as local tissue damage, inflammation and hypotension. Furthermore, the extract neutralized the degradation of the Bbeta chain of human fibrinogen and indirect hemolysis caused by venom. It was also observed that the extract exerted a moderate effect on the clotting time, prolonging it only to a small extent. Edema, hemorrhage and myotoxic effects including lethality, induced by venom were neutralized significantly when different doses of the extract were preincubated with venom before the assays. On the other hand, animals that received extract 10 min after the injection of venom were protected from venom induced toxicity. Since it inhibits hydrolytic enzymes and pharmacological effects, it may be used as an alternative treatment to serum therapy and, in addition, as a rich source of potential inhibitors of PLA(2), metalloproteinases, serine proteases, hyaluronidases and 5 cent-nucleotidases, the enzymes involved in several physiopathological human and animal diseases.
DOI: 10.1002/ptr 866 M. M. MENDES ET AL. properties of the aqueous extract from Pentaclethra macro-loba
- Copyright
Copyright © 2008 John Wiley & Sons, Ltd. Phytother. Res. 22, 859–866 (2008) DOI: 10.1002/ptr 866 M. M. MENDES ET AL. properties of the aqueous extract from Pentaclethra macro-loba. J Ethnopharmacol 100: 145–152.
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Antihemorrhagic, antinucleolytic and other antiophidian
Copyright © 2008 John Wiley & Sons, Ltd.
Phytother. Res. 22, 859-866 (2008)
DOI: 10.1002/ptr