Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology
Inflammatory events induced by brown spider venom and its recombinant dermonecrotic toxin: A pharmacological investigation
Introduction
Brown spiders are found worldwide and spider bites have been reported from all continents (Futrel, 1992, Hogan et al., 2004). Necrotic skin lesions with gravitational spreading and a massive inflammatory response are the hallmark of accidents with the Loxosceles genus (Futrel, 1992, da Silva et al., 2004). Whole venom is composed of a mixture of different proteic toxins, and the mechanisms that lead to these deleterious effects are under investigation.
The dermonecrotic toxin (sphingomyelinase-D, now recognized as phospholipase-D, Lee and Lynch, 2005) is the best known molecule found in different brown spider venoms, and studies have identified it as a contributor to the major toxic effect of spider bites (Binford and Wells, 2003). Previous studies cloned and expressed functional dermonecrotic toxins from Loxosceles intermedia venom glands that were able to reproduce the dermonecrosis and inflammatory response (Chaim et al., 2006, da Silveira et al., 2006, da Silveira et al., 2007).
Recent evidence has shown that the subcutaneous injections of whole L. intermedia venom and the recombinant isoform of the dermonecrotic toxin, LiRecDT1, cause a significant increase in edema and vascular permeability (da Silveira et al., 2006). However, there have been few scientific studies aimed at investigating the mechanisms underlying the brown spider toxin-induced inflammatory actions.
Accumulating data have demonstrated that mast cells are critical contributors to the development of peripheral inflammatory responses associated with tissue or nerve injury (Metcalfe et al., 1997, Mekori and Metcalfe, 2000, Zuo et al., 2003, Liu et al., 2007). Mast cells are intimately involved in the pathophysiology of inflammation (Mekori and Metcalfe, 2000), and their activation promotes the release of chemical mediators responsible for important tissue alterations such as vasodilatation, increases in vascular permeability and neutrophil chemotaxis among others (Metcalfe et al., 1997). Many of these effects are attributable to the activity of histamine and 5-hydroxitryptamine (5-HT, serotonin) of mast cell origin. Some venom phospholipases are known to lead directly to mast cell degranulation, like phospholipase-A (Cirino et al., 1989, Landucci et al., 1998). Additionally, mast cells have phospholipases-D localized in the plasma membrane that are associated to antigen mast cell degranulation (Rivera and Gilfillan, 2006).
The lack of an efficient protocol to alleviate the inflammatory response induced by brown spider envenomation may be related to a reduced number of studies carried out in experimental animals aimed at characterizing the mechanisms involved in inflammatory development. In the present study, we investigated the potential role of mast cells and the involvement of histamine and serotonin in the mechanisms by which the brown spider toxins cause inflammatory events.
Section snippets
Animals
Adult Swiss mice (Mus musculus, 20–30 g), with free access to food and water were used. All protocols employed were in strict accordance with the NIH Guide for the Care and Use of Laboratory Animals and approved by the local ethics committee (process numbers 261/UFPR, 2007). All efforts were made to minimize the number of animals used and their suffering.
Reagents
Compound 48/80, cimetidine, cetirizine, histamine, serotonin and o-phthalaldehyde (OPA) were purchased from Sigma-Aldrich Chemical Co. (St.
Paw edema induced by subcutaneous injections of brown spider toxins
The subcutaneous (s.c.) injection of brown spider whole venom elicited a time (0 to 720 min, 10 µg/paw) (Fig. 1A) and dose-dependent (1 to 20 µg/paw) (Fig. 1B) edema formation that started at 5 min, peaked at 10 min and decreased thereafter, being absent 12 h later (Fig. 1A and B). The s.c. injection of LiRecDT1 also elicited a time (0 to 720 min, 10 µg/paw) (Fig. 1C) and dose-dependent (0.5 to 20 µg/paw) (Fig. 1D) edema formation that peaked at the 10 min time-point and was absent 12 h later (
Discussion
Inflammation is the basic mechanism available for the repair of tissue after an injury and consists of a cascade of cellular and microvascular reactions. This cascade includes edema, elevated permeability in microvessels and the migration of several cell types into damaged tissue (Schmid-Schönbein, 2006). The literature is well-supplied with studies relating the symptoms and signs evoked in accidents caused by brown spiders, but is deficient in studies about the mechanisms underlying the
Acknowledgments
This work was supported by grants from CNPq, CAPES, Fundação Araucária-PR, and Secretaria de Estado de Ciência, Tecnologia e Ensino Superior do Paraná, Brazil.
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2020, International Journal of Biological MacromoleculesCitation Excerpt :Some of these major signals can be observed in Supplementary Fig. S1, which points out the macroscopic appearance of a skin lesion caused by a Brown spider bite in a human injured patient. In addition to the clinical observations in humans, the allergenic potential of Loxosceles venom is supported by animal experiments, which showed after crude venom exposure paw edema, skin edema and increase of vessel permeability [18,42] Moreover, the ability of Loxosceles venom to cause inflammatory reactions was reduced in animals pretreated with compound 48/80, pointing out the involvement of mast cells in these responses. In addition, pretreatment of mice with histamine receptor antagonists, including promethazine and cetirizine, or the serotonin receptor antagonist methysergide significantly decreased venom induced edema and vascular permeability [42].
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2020, ToxiconCitation Excerpt :It is the most well studied and well-characterized group of Loxosceles venom toxins since it is the family of enzymes responsible for its most severe damages. The Loxtox proteins can induce dermonecrosis, strong inflammatory response, platelet aggregation, hemolysis, alteration in vascular permeability, cytotoxicity, nephrotoxicity, acute renal failure, among other clinical symptoms and lethality (Kurpiewski et al., 1981; Rees et al., 1984; Tambourgi et al., 1998; Fernandes-Pedrosa et al., 2002; Kalapothakis et al., 2002; Ramos-Cerrillo et al., 2004; Chaim et al., 2006; Oliveira Christoff et al., 2008; Paludo et al., 2008; Chaves-Moreira et al., 2009; Dias-Lopes et al., 2010; Chaves-Moreira et al., 2011; Gomes et al., 2011; Gremski et al., 2014). The name Loxtox was proposed by Kalapothakis et al. (2007) as a broader designation for the set of homologous genes from Loxosceles, since some PLDs expressed in the venom show undetectable SMase D or PLD activity.
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2016, Biochimica et Biophysica Acta - Molecular and Cell Biology of LipidsCitation Excerpt :Previous studies have reported that enzymatic hydrolysis of membrane phospholipids is necessary for the biological activities triggered by these PLD toxins. Phospholipase D homologs devoid of catalytic activity are unable to induce dermonecrotic lesions in rabbits [17,24–28,37]. Moreover, toxins with low or residual enzymatic activity on synthetic sphingomyelin exhibit reduced or residual hemolytic activity on human erythrocytes and weaker or residual effects on platelet aggregation and vascular permeability [17,24–28,37].