ABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Snakebite envenoming is a public health problem of high impact in Central America. Bothrops asper, known as barba amarilla, terciopelo, and equis, is the snake species responsible for most snakebites in Central America. In this region, there is a long-standing tradition on the use of plants in the management of snakebites, especially in indigenous communities. Ethnomedical use of Eryngium foetidum L., Neurolaena lobata (L.) Cass. and Pimenta dioica (L.) Merr. to treat snakebite envenoming has been reported in Belice, Guatemala, Nicaragua, and Costa Rica. Extracts of the leaves of these plants have shown anti-venom activities in in vitro assays in previous studies. AIM OF THE STUDY: To assess the ability of organic fractions from these three plants to inhibit enzymatic activities associated with toxicity of the venom of B. asper, and to study, by docking analysis, the interaction of metalloproteinase and phospholipases A2 (PLA2) from B. asper venom with secondary metabolites previously described in these plants. MATERIALS AND METHODS: Organic fractions were obtained from these three plant species and their ability to neutralize proteolytic, PLA2 and in vitro coagulant activities of B. asper venom was assessed. A phytochemical analysis was carried out in these fractions. The interaction of secondary metabolites previously described in these plants with three toxins from B. asper venom (a metalloproteinase, a PLA2 and a PLA2 homologue) was investigated by docking analysis. RESULTS: The inhibitory activity of plants was mainly concentrated in their polar fractions. Acetonic fraction from P. dioica was the most active against PLA2 activity, while the acetonic fraction of E. foetidum completely inhibited the proteolytic activity of the venom. Coagulant activity was partially inhibited only by the acetone and ethyl acetate fractions of P. dioica. Phytochemical analysis of the most bioactive fractions identified flavonoids, saponins, essential oils, coumarins, alkaloids, tannins and sesquiterpene lactones. Docking analysis revealed high affinity interactions of several secondary metabolites of these plants with residues in the vicinity of the catalytic site of these enzymes and, in the case of PLA2 homologue myotoxin II, in the hydrophobic channel. CONCLUSIONS: Various fractions from these plants have inhibitory activity against enzymatic actions of B. asper venom which are directly associated with toxicological effects. Docking analysis showed structural evidence of the interaction of secondary metabolites with three toxins. These observations provide support to the potential of these plants to inhibit relevant toxic components of this snake venom.
Subject(s)
Antivenins/pharmacology , Crotalid Venoms/antagonists & inhibitors , Plant Extracts/pharmacology , Snake Bites/drug therapy , Animals , Antivenins/isolation & purification , Asteraceae/chemistry , Bothrops , Central America , Eryngium/chemistry , Humans , Medicine, Traditional , Molecular Docking Simulation , Phytochemicals/chemistry , Phytochemicals/isolation & purification , Phytochemicals/pharmacology , Pimenta/chemistry , Plant LeavesABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Snake bite is a major occupational hazard in tropical and subtropical countries including India as per the World Health Organization. Naja naja (Indian cobra) and Daboia russelli (Russell's viper) are the two poisonous snakes commonly associated with human mortality in India. Andrographis serpyllifolia (Rottler ex Vahl) Wight has been documented in ethnobotanical records as a plant possessing potent anti-snake venom activity. AIM OF THE STUDY: The present study is aimed for systematic evaluation of in vitro anti-venom potential of various solvent based leaf extracts of A. serpyllifolia against toxic venom enzymes of Naja naja and Daboia russelli. MATERIALS AND METHODS: Different solvent based leaf extracts of A. serpyllifolia were tested against the snake venoms of Naja naja and Daboia russelli obtained from Irula Snake Catchers Industrial Co-operative Society Limited, Kancheepuram, Tamil nadu, India. Three different in vitro neutralization assays such as indirect hemolysis, procoagulent and lytic activities and seven in vitro enzyme inhibition assays such as protease, acetylcholinesterase, phosphomonoesterase, phosphodiesterase, 5'nucleotidase, phospholipase A2, hyaluronidase and post synaptic acetylcholine receptor binding activity were carried out according to standard protocols. The results were analyzed using the standard ANOVA procedures. RESULTS: Among various solvent based leaf extracts of A. serpyllifolia tested, aqueous extract showed maximum neutralizing and inhibitory activities against Naja naja and Daboia russelli venoms. CONCLUSIONS: The various in vitro enzymatic studies reveal that the aqueous leaf extract of A. serpyllifolia plant could inhibit most of the toxic enzymes of the Naja naja and Daboia russelli venoms which could be further confirmed by in vivo studies.
Subject(s)
Andrographis , Antivenins/pharmacology , Elapid Venoms/antagonists & inhibitors , Plant Extracts/pharmacology , Solvents/pharmacology , Viper Venoms/antagonists & inhibitors , Animals , Antivenins/isolation & purification , Dose-Response Relationship, Drug , Elapid Venoms/isolation & purification , Naja naja , Plant Extracts/isolation & purification , Plant Leaves , Solvents/isolation & purification , Viper Venoms/isolation & purificationABSTRACT
BACKGROUND: In Brazil, the Bothrops genus accounts for 87% of registered snakebites, which are characterized by hemorrhage, tissue necrosis, hemostatic disturbances, and death. The treatment recommended by governments is the administration of specific antivenoms. Although antivenom efficiently prevents venom-induced lethality, it has limited efficacy in terms of preventing local tissue damage. Thus, researchers are seeking alternative therapies able to inhibit the main toxic effects of venoms, without compromising safety. OBJECTIVE: The study aimed to test the ability of aqueous extracts of leaves, stems, and fruits of the plant Clusia fluminensis to neutralize some toxic effects induced by the venoms of Bothrops jararaca and Bothrops jararacussu. METHODS: The plant extracts were incubated with venoms for 30 min. at 25 °C, and then in vitro (coagulant and proteolytic) and in vivo (hemorrhagic, myotoxic, and edematogenic) activities were evaluated. In addition, the extracts were administered to animals (by oral, intravenous or subcutaneous routes) before or after the injection of venom samples, and then hemorrhage and edema assays were performed. In addition, a gel solution of the fruit extract was produced and tested in terms of reducing hemorrhage effects. A chemical prospection was performed to identify the main classes of compounds present in the extracts. RESULTS: All the extracts inhibited the activities of the two venoms, regardless of the experimental protocol or route of administration of the extracts. Moreover, the gel of the fruit extract inhibited the venom-induced-hemorrhage. The extracts comprised of tannins, flavonoids, saponins, steroids, and terpenoids. CONCLUSION: Antivenom properties of C. fluminensis extracts deserve further investigation in order to gain detailed knowledge regarding the neutralization profile of these extracts.
Subject(s)
Antivenins/pharmacology , Clusia/chemistry , Plant Extracts/pharmacology , Snake Venoms/antagonists & inhibitors , Animals , Antivenins/chemistry , Antivenins/isolation & purification , Bothrops , Brazil , Fruit/chemistry , Hemorrhage/drug therapy , Mice , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Plant Leaves/chemistry , Plant Stems/chemistry , Snake Venoms/toxicityABSTRACT
Snakebite envenomation is an important health problem in tropical countries, with severe human and social consequences. In Latin America, the Bothrops species constitute the main threat to humans, and the envenomation caused by these species quickly develops 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 which are 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.
Subject(s)
Antivenins/pharmacology , Biological Products/pharmacology , Phytochemicals/pharmacology , Plant Extracts/pharmacology , Snake Venoms/antagonists & inhibitors , Animals , Antivenins/chemistry , Antivenins/isolation & purification , Biological Products/chemistry , Biological Products/isolation & purification , Bothrops , Humans , Molecular Structure , Phytochemicals/chemistry , Phytochemicals/isolation & purification , Plant Extracts/chemistry , Plant Extracts/isolation & purificationABSTRACT
Bredemeyera floribunda roots are popularly used to treat snakebites in the semiarid region of Northeast Brazil, and previous studies indicate the anti-ophidian actions of triterpenoid saponins found in its roots. To assess B. floribunda root extract (BFRE) activity against the effects of Bothrops jararacussu venom (BjuV), antiphospholipasic, antiproteolytic, antihemorrhagic, antinecrotic, and anti-edematogenic activities were investigated in mice. Phytochemical analysis revealed the presence of saponins, flavonoids, and sugars, with rutin and saccharose being the major constituents of BFRE. Acute toxicity was determined and BFRE was nontoxic to mice. Phospholipase A2 and proteolytic activities induced by BjuV were inhibited in vitro by BFRE at all concentrations tested herein. BFRE (150 mg/kg) inhibited paw edema induced by BjuV (50 µg/animal), reducing total edema calculated by area under the curve, but carrageenan-induced paw edema was unchanged. Hemorrhagic and necrotizing actions of BjuV (50 µg/animal) were considerably decreased by BFRE treatment. Thus, BFRE blocked the toxic actions of B. jararacussu venom despite having no anti-inflammatory activity, which points to a direct inhibition of venom's toxins, as demonstrated in the in vitro assays. The larger amounts of rutin found in BFRE may play a role in this inhibition, since 3′,4′-OH flavonoids are known inhibitors of phospholipases A2.
Subject(s)
Animals , Male , Rats , Antivenins/pharmacology , Plant Extracts/pharmacology , Plant Roots/chemistry , Crotalid Venoms/antagonists & inhibitors , Edema/drug therapy , Hemorrhage/etiology , Antivenins/isolation & purification , Bothrops , Crotalid Venoms/toxicity , Polygalaceae/chemistry , Disease Models, Animal , Dose-Response Relationship, Drug , Edema/etiology , Hemorrhage/drug therapyABSTRACT
Bredemeyera floribunda roots are popularly used to treat snakebites in the semiarid region of Northeast Brazil, and previous studies indicate the anti-ophidian actions of triterpenoid saponins found in its roots. To assess B. floribunda root extract (BFRE) activity against the effects of Bothrops jararacussu venom (BjuV), antiphospholipasic, antiproteolytic, antihemorrhagic, antinecrotic, and anti-edematogenic activities were investigated in mice. Phytochemical analysis revealed the presence of saponins, flavonoids, and sugars, with rutin and saccharose being the major constituents of BFRE. Acute toxicity was determined and BFRE was nontoxic to mice. Phospholipase A2 and proteolytic activities induced by BjuV were inhibited in vitro by BFRE at all concentrations tested herein. BFRE (150 mg/kg) inhibited paw edema induced by BjuV (50 µg/animal), reducing total edema calculated by area under the curve, but carrageenan-induced paw edema was unchanged. Hemorrhagic and necrotizing actions of BjuV (50 µg/animal) were considerably decreased by BFRE treatment. Thus, BFRE blocked the toxic actions of B. jararacussu venom despite having no anti-inflammatory activity, which points to a direct inhibition of venom's toxins, as demonstrated in the in vitro assays. The larger amounts of rutin found in BFRE may play a role in this inhibition, since 3',4'-OH flavonoids are known inhibitors of phospholipases A2.
Subject(s)
Antivenins/pharmacology , Crotalid Venoms/antagonists & inhibitors , Edema/drug therapy , Hemorrhage/etiology , Plant Extracts/pharmacology , Plant Roots/chemistry , Polygalaceae/chemistry , Animals , Antivenins/isolation & purification , Bothrops , Crotalid Venoms/toxicity , Disease Models, Animal , Dose-Response Relationship, Drug , Edema/etiology , Hemorrhage/drug therapy , Male , RatsABSTRACT
Nowadays, treatment with specific antivenins is considered the only cure for snakebites accidents. However, access to antivenom obstructs the successful implementation of the World Health Organization international guidelines. In the last few years, natural organic compounds, peptides, and proteins with the ability to inhibit snake toxins and obtained from different sources such as plant extracts and animal blood have been proposed as antivenoms. In this work, we will focus on the inhibitors of the main venom toxins, phospholipases A2 and metalloproteinases, and their application as novel antivenoms.
Subject(s)
Antivenins/pharmacology , Biological Products/pharmacology , Snake Venoms/antagonists & inhibitors , Animals , Antivenins/chemistry , Antivenins/isolation & purification , Biological Products/chemistry , Biological Products/isolation & purification , HumansABSTRACT
This is the first report on large-scale experimental production of an equine antivenom against the redback spider (Latrodectus hasseltii) lived in Japan. We captured 10,000 redback spiders in Japan and prepared the toxoids of crude venom extract, mixed the toxoids with a mineral oil adjuvant, and immunized healthy horses repeatedly over a period of several weeks. Thereafter, we separated the horse plasma, purified the γ-globulin fraction, and stocked it as a purified antivenom concentrate. Consequently, we manufactured approximately 6,500 vials of a single-dose freeze-dried test lot from a portion of the purified γ-globulin fraction, equivalent to the extract derived from 520 spiders. This test lot had an antitoxin titer comparable to that of a similar drug commercially available overseas (a liquid preparation), and the other quality met all quality reference specifications based on the Minimum Requirements for Biological Products and other guidelines relevant to existing antivenom drug products in Japan.
Subject(s)
Antivenins , Spiders/drug effects , Venoms , Animals , Antigens/immunology , Antivenins/biosynthesis , Antivenins/immunology , Antivenins/isolation & purification , Horses , Immunization , Spiders/immunology , Venoms/immunologyABSTRACT
OBJECTIVES: Snakebites are a significant and severe global health problem. Till date, anti-snake venom serum is the only beneficial remedy existing on treating the snakebite victims. As antivenom was reported to induce early or late adverse reactions to human beings, snake venom neutralizing potential for Cyclea peltata root extract was tested for the present research by ex vivo and in vivo approaches on Naja naja toxin. MATERIALS AND METHODS: Ex vivo evaluation of venom toxicity and neutralization assays was carried out. The root extracts from C. peltata were used to evaluate the Ex vivo neutralization tests such as acetylcholinesterase, protease, direct hemolysis assay, phospholipase activity, and procoagulant activity. Gas chromatography-mass spectrometry (GC-MS) analysis from root extracts of C. peltata was done to investigate the bioactive compounds. RESULTS: The in vivo calculation of venom toxicity (LD50) of N. naja venom remained to be 0.301 µg. C. peltata root extracts were efficiently deactivated the venom lethality, and effective dose (ED50) remained to be 7.24 mg/3LD50 of N. naja venom. C. peltata root extract was found effective in counteracting all the lethal effects of venom. GC-MS analysis of the plant extract revealed the presence of antivenom compounds such as tetradecanoic and octadecadienoic acid which have neutralizing properties on N. naja venom. CONCLUSION: The result from the ex vivo and in vivo analysis indicates that C. peltata plant root extract possesses significant compounds such as tetradecanoic acid hexadecanoic acid, heptadecanoic acid, and octadecadienoic acid which can counteract the toxins present in N. naja.
Subject(s)
Antivenins/therapeutic use , Cyclea , Elapid Venoms/antagonists & inhibitors , Plant Extracts/therapeutic use , Plant Roots , Snake Bites/drug therapy , Acetylcholinesterase/metabolism , Animals , Antivenins/isolation & purification , Antivenins/pharmacology , Dose-Response Relationship, Drug , Hemolysis/drug effects , Hemolysis/physiology , Mice , Naja , Plant Extracts/isolation & purification , Plant Extracts/pharmacology , Snake Bites/enzymologyABSTRACT
Snakebite is a serious occupational hazard affecting mainly rural populations of tropical and subtropical developing countries. Lachesis muta (Bushmaster) bites are extremely serious but are rarely reported in the literature. Bushmaster envenomings are characterized by intense local pain, edema, neurotoxicity, hypotension, local hemorrhage, and dramatic systemic alterations. Antivenom treatment has regularly been used for more than a century; however, it fails to neutralize local tissue damage and hemorrhage, leading to morbidity or disabilities in victims. Thus, the production and clinical use of antivenom must be improved. The present work characterizes, for the first time, a sulfated polysaccharide from the red seaweed, Laurencia aldingensis, including its neutralizing effect on some toxic activities of L. muta venom. Chemical and spectroscopic analyses showed that L. aldingensis produces sulfated agarans with the A-units partially C-2 sulfated or 6-O-methoxylated presetting the B-units in the cyclized (3,6-anhydro-α-L-galactose) or in the non-cyclized form (α-L-galactose). The latter is significantly substituted by sulfate groups on C-6. In vitro and in vivo assays showed that this sulfated agaran inhibited hemolysis, coagulation, proteolysis, edema, and hemorrhage of L. muta venom. Neutralization of hemorrhagic activity was also observed when the agaran was administered by different routes and after or before the venom injection. Furthermore, the agaran blocked the edema caused by a phospholipase A2 isolated from the L. muta venom. Experimental evidence therefore indicates that the sulfated agaran of L. aldingensis has potential to aid antivenom therapy of accidents caused by L. muta venom and may help to develop more effective antivenom treatments of snake bites in general.
Subject(s)
Antivenins/pharmacology , Edema/prevention & control , Laurencia/chemistry , Polysaccharides/pharmacology , Snake Bites/drug therapy , Viper Venoms/antagonists & inhibitors , Animals , Antivenins/chemistry , Antivenins/isolation & purification , Blood Coagulation/drug effects , Edema/chemically induced , Hemolysis/drug effects , Hemorrhage/chemically induced , Hemorrhage/prevention & control , Humans , Mice , Phospholipases A2/administration & dosage , Plant Extracts/chemistry , Polysaccharides/chemistry , Polysaccharides/isolation & purification , Proteolysis/drug effects , Seaweed , Snake Bites/physiopathology , Sulfates , Viper Venoms/toxicity , ViperidaeABSTRACT
In our earlier study, we have reported that a phenolic compound 2-hydroxy-4-methoxybenzaldehyde from Janakia arayalpatra root extract was active against Viper and Cobra envenomations. Based on the structure of this natural product, libraries of synthetic structurally variant phenolic compounds were studied through molecular docking on the venom protein. To validate the activity of eight selected compounds, we have tested them in in vivo and in vitro models. The compound 21 (2-hydroxy-3-methoxy benzaldehyde), 22 (2-hydroxy-4-methoxybenzaldehyde) and 35 (2-hydroxy-3-methoxybenzylalcohol) were found to be active against venom-induced pathophysiological changes. The compounds 20, 15 and 35 displayed maximum anti-hemorrhagic, anti-lethal and PLA2 inhibitory activity respectively. In terms of SAR, the presence of a formyl group in conjunction with a phenolic group was seen as a significant contributor towards increasing the antivenom activity. The above observations confirmed the anti-venom activity of the phenolic compounds which needs to be further investigated for the development of new anti-snake venom leads.
Subject(s)
Antivenins/chemistry , Antivenins/pharmacology , Biological Products/pharmacology , Models, Molecular , Phenols/pharmacology , Phospholipase A2 Inhibitors/pharmacology , Phospholipases A2/metabolism , Snake Venoms/enzymology , Antivenins/isolation & purification , Biological Products/chemistry , Biological Products/isolation & purification , Dose-Response Relationship, Drug , Molecular Structure , Phenols/chemistry , Phenols/isolation & purification , Phospholipase A2 Inhibitors/chemistry , Phospholipase A2 Inhibitors/isolation & purification , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Plant Extracts/pharmacology , Plant Roots/chemistry , Structure-Activity RelationshipABSTRACT
BACKGROUND: Every year thousands of people are victims of burns, mainly scald burns. Many of these victims have small size wounds and superficial partial thickness and do not seek specialized medical care. As in Brazil Casearia sylvestris Sw., popularly known as guaçatonga is widely used for its analgesic, antiseptic and anti-inflammatory activities, this study sought to evaluate the effects of its hydroalcoholic extract in healing process of burns injuries. METHODS: The obtained extract was validated applying a thin layer chromatography and sophisticated validation method using Bothrops jararacussu snake venom that is necrotic and inflammatory, and by which guaçatonga extract was able to neutralize the irreversible neuromuscular blockade induced by the venom. After induction of the scald injury, the animals were treated daily with saline solution spray; spray containing extract; biofilm; or biofilm impregnated with extract. RESULTS: Significant differences were observed between the four groups studied considering: extension of the healing area, neovascularization, fibroblast proliferation, and epithelialization. CONCLUSION: The anti-inflammatory and bactericidal effects of C. sylvestris Sw. suggests a potential therapeutic benefit in the treatment of inflammatory conditions in second-degree scald burn injuries, as well as, counteracting against the in vitro paralysis induced by B. jararacussu venom.
Subject(s)
Analgesics/pharmacology , Anti-Inflammatory Agents/pharmacology , Antivenins/pharmacology , Burns/drug therapy , Casearia/chemistry , Neuroprotective Agents/pharmacology , Wound Healing/drug effects , Analgesics/isolation & purification , Animals , Anti-Inflammatory Agents/isolation & purification , Antivenins/isolation & purification , Bothrops/metabolism , Burns/pathology , Crotalid Venoms/antagonists & inhibitors , Crotalid Venoms/isolation & purification , Crotalid Venoms/toxicity , Hot Temperature , Male , Mice , Neuroprotective Agents/isolation & purification , Plant Extracts/chemistry , Rats, Wistar , Skin/drug effects , Skin/injuries , Skin/pathologyABSTRACT
In Brazil, snakebites are a public health problem and accidents caused by Lachesis muta have the highest mortality index. Envenomation by L. muta is characterized by systemic (hypotension, bleeding and renal failure) and local effects (necrosis, pain and edema). The treatment to reverse the evolution of all the toxic effects is performed by injection of antivenom. However, such therapy does not effectively neutralize tissue damage or any other local effect, since in most cases victims delay seeking appropriate medical care. In this way, alternative therapies are in demand, and molecules from natural sources have been exhaustively tested. In this paper, we analyzed the inhibitory effect of a sulfated galactan obtained from the red seaweed Palisada flagellifera against some toxic activities of L. muta venom. Incubation of sulfated galactan with venom resulted in inhibition of hemolysis, coagulation, proteolysis, edema and hemorrhage. Neutralization of hemorrhage was also observed when the galactan was administered after or before the venom injection; thus mimicking a real in vivo situation. Moreover, the galactan blocked the edema caused by a phospholipase A2 isolated from the same venom. Therefore, the galactan from P. flagellifera may represent a promising tool to treat envenomation by L. muta as a coadjuvant for the conventional antivenom.
Subject(s)
Antivenins/pharmacology , Galactans/pharmacology , Rhodophyta/chemistry , Viper Venoms/antagonists & inhibitors , Animals , Antivenins/isolation & purification , Brazil , Galactans/isolation & purification , Mice , Mice, Inbred BALB C , Phospholipases A2/metabolism , Snake Bites/drug therapy , Viper Venoms/toxicity , ViperidaeABSTRACT
ETHNOPHARMACOLOGY RELEVANCE: In Colombia, more than 4.000 ophidian accidents occur per year and due to the scarce distribution and limited availability of antivenom, the use of traditional medicine has been perpetuated in some of its rural communities, in which initially, those affected are treated by healers and shamans using medicinal plants in different ways. METHODS: Research was conducted with renowned healers or connoisseurs of plants on the ethnobotany of ophidian accidents in five different areas and their municipalities of Antioquia: Magdalena Medio (Caracolí, Puerto Berrío); Bajo Cauca (Caucasia, Zaragoza); Nordeste (San Roque, Yalí); Norte (Gómez Plata, Valdivia); Suroeste (Ciudad Bolívar, Salgar); collecting information related to experience and time of use of plants in the treatment of these poisonings, amounts used, ways of use (beverage, bathing, ointment, chupaderas, vapors), preparation types (maceration or decoction) and treatment duration. RESULTS: 71 plant species were identified and collected, 49.29% of them without previous reports as antiophidian and 38.0% employed for the same purpose in other geographical areas. The leaves (24.82%), stems (11.68%) and flowers (10.95%) were found to be the most frequently employed structures in the preparation of the extracts, which are usually prepared by decoction (83.94%), maceration (6.57%). CONCLUSIONS: In this work, specimens lacking previous ethnobotanical reports have been found, plants used by ethnic groups from other regions of Antioquia and the world to treat snake bites; and herbaceous plants whose inhibitory activity of symptoms produced by some snake venoms, has been experimentally verified by in vivo and in vitro tests.
Subject(s)
Antivenins/therapeutic use , Plant Preparations/therapeutic use , Plants, Medicinal/chemistry , Snake Bites/drug therapy , Adult , Aged , Aged, 80 and over , Antivenins/isolation & purification , Colombia , Ethnopharmacology , Health Knowledge, Attitudes, Practice , Humans , Medicine, Traditional , Middle Aged , Phytotherapy/methods , Plant Preparations/isolation & purificationABSTRACT
The aqueous extract of Mangifera indica is known to possess anti-snake venom activities. However, its inhibitory potency and mechanism of action on multi-toxic phospholipases A2s, which are the most toxic and lethal component of snake venom is still unknown. Therefore, this study was carried out to evaluate the modulatory effect of standard aqueous bark extract of M. indica on VRV-PL-VIIIa of Indian Russells viper venom. Mangifera indica extract dose dependently inhibited the GIIB sPLA2 (VRV-PL-VIIIa) activity with an IC50 value of 6.8±0.3 µg/ml. M. indica extract effectively inhibited the indirect hemolytic activity up to 96% at ~40 µg/ml concentration. Further, M. indica extract at different concentrations (0-50 µg/ml) inhibited the edema formed in a dose dependent manner. It was found that there was no relieve of inhibitory effect of the extract when examined as a function of increased substrate and calcium concentration. The inhibition was irreversible as evident from binding studies. The in vitro inhibition is well correlated with in situ and in vivo edema inducing activities. As the inhibition is independent of substrate, calcium concentration and was irreversible, it can be concluded that M. indica extracts mode of inhibition could be due to direct interaction of components present in the extract with PLA2 enzyme. In conclusion, the aqueous extract of M. indica effectively inhibits svPLA2 (Snake venom phospholipase A2) enzymatic and its associated toxic activities, which substantiate its anti-snake venom properties. Further in-depth studies are interesting to known on the role and mechanism of the principal inhibitory constituents present in the extract, so as to develop them into potent anti-snake venom and as an anti-inflammatory agent.
Subject(s)
Antivenins/metabolism , Group II Phospholipases A2/antagonists & inhibitors , Mangifera/chemistry , Plant Bark/chemistry , Plant Extracts/metabolism , Animals , Antivenins/isolation & purification , Disease Models, Animal , Dose-Response Relationship, Drug , Erythrocytes/drug effects , Hemolysis/drug effects , Humans , Inhibitory Concentration 50 , Mice , Plant Extracts/isolation & purification , Poisoning/prevention & controlABSTRACT
Significant inhibition of the coagulant and hemorrhagic effects of Bothrops asper venom was demonstrated by ethanolic extract prepared from the leaves of Brownea rosademonte. In vitro experiments preincubating 5.5 mg of extract kg-1 b.m. for 30 min with a minimum hemorrhagic dose of venom (273.8 ± 16.1 µg of venom kg-1 b.m.) lowered the hemorrhagic activity of the venom alone in CD-1 mice by 51.5 ± 2.6 %. Additionally, 1.7 mg extract L-1 plasma prolonged 5.1 times the plasma coagulation time. Fractionation of the extract led to the isolation of two compounds: ononitol (1) and quercetrin (2). The structure of compounds 1 and 2 was established by spectroscopic analyses, including APCI-HRMS and NMR (1H, 13C, HSQC, HMBC and COSY). A quercetrin concentration of 0.11 µmol L-1 prolonged the plasma coagulation time 2.6 times demonstrating that this compound was one of the active constituents of the Brownea rosademonte extract.
Subject(s)
Antivenins/pharmacology , Crotalid Venoms/antagonists & inhibitors , Fabaceae/chemistry , Plant Extracts/pharmacology , Animals , Antivenins/isolation & purification , Blood Coagulation/drug effects , Bothrops , Ethanol/chemistry , Glycosides/isolation & purification , Glycosides/pharmacology , Hemorrhage/prevention & control , Mice , Plant Extracts/chemistry , Plant Leaves , Quercetin/analogs & derivatives , Quercetin/isolation & purification , Quercetin/pharmacology , Spectrum AnalysisABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Various medicinal plants have protective properties against the toxicities of the venom of cobra snake (Naja species). They may be used as local first aid for the treatment of snakebite victims, and can significantly inhibit lethality, cardio-, neuro-, nephro- and myotoxicity, hemorrhage, and respiratory paralysis induced by the cobra snake venom. The plants or their extracts may also complement the benefits of conventional anti-serum treatment. AIM OF THE REVIEW: This review provides information on the protective, anti-venom, properties of medicinal plants against snakebites from cobras. In addition, it identifies knowledge gaps and suggests further research opportunities. METHODS: The literature was searched using databases including Google Scholar, PubMed, ScienceDirect, Scopus and Web of Science. The searches were limited to peer-reviewed journals written in English with the exception of some books and a few articles in foreign languages. RESULTS: The plants possess neutralization properties against different cobra venom enzymes, such as hyaluronidase, acetylcholinesterase, phospholipase A2 and plasma proteases. Different active constituents that show promising activity against the effects of cobra venom include lupeol acetate, ß-sitosterol, stigmasterol, rediocides A and G, quercertin, aristolochic acid, and curcumin, as well as the broad chemical groups of tannins, glycoproteins, and flavones. The medicinal plants can increase snakebite victim survival time, decrease the severity of toxic signs, enhance diaphragm muscle contraction, block antibody attachment to venom, and inhibit protein destruction. In particular, the cardiovascular system is protected, with inhibition of blood pressure decline and depressed atrial contractility and rate, and prevention of damage to heart and vessels. The designs of experimental studies that show benefits, or otherwise, of use of medicinal plants have some limitations: deficiency in identification and isolation of active constituents responsible for therapeutic activity; lack of comparison with reference drugs; and little investigation of the mechanism of anti-venom activity. CONCLUSION: Despite some current deficiencies in experimental or clinical analysis, medicinal plants with anti-venom characteristics are effective and so are candidates for future therapeutic agents. We suggest that emphasis on identification and testing of active ingredients in research in the future will assist better understanding of their anti-venom activity.
Subject(s)
Antivenins/pharmacology , Plant Preparations/pharmacology , Plants, Medicinal/chemistry , Animals , Antivenins/isolation & purification , Elapid Venoms/antagonists & inhibitors , Elapidae , Humans , Snake Bites/drug therapyABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: The high frequency of poisoning by sting or bite from venomous animals has begun to be a serious public health problem in Mexico where scorpion sting is the most common. Because of this, there is the need to seek active substances in plant species with an antagonistic effect against neurotropic activity of scorpion venom. The aim of this work was to demonstrate which of the compounds contained in the n-hexane extract from Aristolochia elegans roots display activity against scorpion venom. MATERIAL AND METHODS: Antagonist activity displayed by extract, fractions and isolated compounds obtained from Aristolochia elegans was guided by the inhibition of smooth muscle contraction induced by scorpion venom (Centruroides limpidus limpidus) in a model of isolated guinea pig ileum. The neolignans obtained from this extract were isolated and analyzed by chromatographic methods including HPLC. The chemical characterization of these compounds was performed by the analysis of (1)H and (13)C NMR spectra. RESULTS: The bio-guided chromatographic fractionation allowed us to isolate 4 known neolignans: Eupomatenoid-7 (1), licarin A (2), licarin B (3), eupomatenoid-1 (4) and other new neolignan which was characterized as 2-(3'-hydroxy-4'-methoxyphenyl)-3-methyl-5-[(E)-α-propen-γ-al]-7-methoxy-benzo [b] furan (5). This compound was named as eleganal. Compounds 1 and 2 were purified from the most active fraction AeF3 (EC50 of 149.9µg/mL, Emax of 65.66%). A doses-response analysis of eupomatenoid-7(1) and licarin A(2) allowed us to establish EC50 values (65.96µg/mL and 51.96µg/mL) respectively. CONCLUSIONS: The antagonistic effect against Centuroides limpidus limpidus scorpion venom displayed by the n-hexane extract from Aristolochia elegans roots is due to the presence of neolignans 1-2 contained in the fraction AeF3. Chemical analysis of fraction AeF2 allowed the isolation of a new compound which was identified as 2-(3'-hydroxy-4'-methoxyphenyl)-3-methyl-5-[(E)-α-propen-γ-al]-7-methoxy-benzo[b]furan (5), denominated as eleganal.
Subject(s)
Antivenins/pharmacology , Aristolochia/chemistry , Lignans/pharmacology , Scorpion Venoms/antagonists & inhibitors , Animals , Antivenins/administration & dosage , Antivenins/isolation & purification , Bites and Stings/drug therapy , Chromatography, High Pressure Liquid/methods , Dose-Response Relationship, Drug , Guinea Pigs , Lignans/administration & dosage , Lignans/isolation & purification , Magnetic Resonance Spectroscopy , Male , Mexico , Muscle Contraction/drug effects , Muscle, Smooth/drug effects , Muscle, Smooth/metabolism , Plant Extracts/administration & dosage , Plant Extracts/pharmacology , Plant Roots , ScorpionsABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Snakebite envenomation, every year, causes estimated 5-10,000 mortalities and results in more than 5-15,000 amputations in sub-Saharan Africa alone. Antiserum is not easily accessible in these regions or doctors are simply not available, thus more than 80% of all patients seek traditional practitioners as first-choice. Therefore it is important to investigate whether the plants used in traditional medicine systems contain compounds against the necrosis-inducing enzymes of snake venom. MATERIALS AND METHODS: Extracts from traditionally used plants from DR Congo, Mali and South Africa were tested in hyaluronidase, phospholipase A2 and protease enzyme bioassays using Bitis arietans and Naja nigricollis as enzyme source. RESULTS: A total of 226 extracts from 94 different plant species from the three countries, Mali, Democratic Republic of Congo and South Africa were tested in phospholipase A2, proteases and hyaluronidase enzyme assays. Forty plant species showed more than 90% inhibition in one or more assay. Fabaceae, Anacardiaceae and Malvaceae were the families with the highest number of active species, and the active compounds were distributed in different plant parts depending on plant species. Polyphenols were removed in the search for specific enzyme inhibitors against hyaluronidase, phospholipase A2 or proteases from extracts with IC50 values below 100µg/ml. Water extracts of Pupalia lappacea, Combretum molle, Strychnos innocua and Grewia mollis and ethanol extract of Lannea acida and Bauhinia thonningii still showed IC50 values below 100µg/ml in either the hyaluronidase or protease bioassay after removal of polyphenols. CONCLUSION: As four of the active plants are widely distributed in the areas where the snake species Bitis arietans and Naja nigricollis occur a potential inhibitor of the necrotic enzymes is accessible for many people in sub-Saharan Africa.
Subject(s)
Antivenins/pharmacology , Plant Extracts/pharmacology , Plants, Medicinal/chemistry , Snake Venoms/antagonists & inhibitors , Antivenins/administration & dosage , Antivenins/isolation & purification , Bites and Stings/drug therapy , Democratic Republic of the Congo , Hyaluronoglucosaminidase/antagonists & inhibitors , Inhibitory Concentration 50 , Mali , Medicine, African Traditional , Necrosis , Peptide Hydrolases/drug effects , Phospholipases A2/drug effects , Plant Extracts/administration & dosage , Snake Venoms/enzymology , South AfricaABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Serotherapy against snakebite is often unavailable in some regions over Brazil, where people make use of plants from folk medicine to deal with ophidic accidents. About 10% of Combretum species have some ethnopharmacological use, including treatment of snakebites. MATERIALS AND METHODS: We evaluated the ability of the extract of Combretum leprosum and its component arjunolic acid to reduce some in vivo and in vitro effects of Bothrops jararacussu and Bothrops jararaca venoms. The protocols investigated include phospholipase, proteolytic, collagenase, hyaluronidase, procoagulant, hemorrhagic, edematogenic, myotoxic and lethal activities induced by these venoms in Swiss mice. RESULTS: Oral pre-treatment with arjunolic acid reduced the Bothrops jararacussu lethality in up to 75%, while preincubation prevented the death of all the animals. Hemoconcentration effect of Bothrops jararacussu venom was confirmed two hours after i.p. injection, while preincubation with arjunolic acid preserved the hematocrit levels. Both Combretum leprosum extract and arjunolic acid abolished the myotoxic action of Bothrops jararacussu venom. Preincubation of Bothrops jararacussu venom with the extract or arjunolic acid prevented the increase of plasma creatine kinase activity in mice. The hemorrhagic activity of Bothrops jararaca crude venom was reduced down to about 90% and completely inhibited by preincubation with 10 mg/kg or 100 mg/kg Combretum leprosum extract, respectively, while the preincubation and the pretreatment with 30 mg/kg of arjunolic acid reduced the venom hemorrhagic activity down to about 12% and 58%, respectively. The preincubation of the venom with both extract and 30 mg/kg arjunolic acid significantly reduced the bleeding amount induced by Bothrops jararacussu venom. The extract of Combretum leprosum decreased the edema formation induced by Bothrops jararacussu venom both in preincubation and pretreatment, but not in posttreatment. Similarly, arjunolic acid preincubated with the venom abolished edema formation, while pre- and posttreatment have been partially effective. Some enzymatic activities of Bothrops jararacussu and Bothrops jararaca venoms, i.e. phospholipase A2, collagenase, proteolytic and hyaluronidase activities, were to some extent inhibited by the extract and arjunolic acid in a concentration-dependent manner. CONCLUSIONS: Altogether, our results show that Combretum leprosum extract can inhibit different activities of two important Brazilian snake venoms, giving support for its popular use in folk medicine in the management of venomous snakebites.