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1.
Toxins (Basel) ; 15(11)2023 10 25.
Artículo en Inglés | MEDLINE | ID: mdl-37999488

RESUMEN

L-Amino acid oxidase (LAAO) is an enzyme found in snake venom that has multifaceted effects, including the generation of hydrogen peroxide (H2O2) during oxidative reactions, leading to various biological and pharmacological outcomes such as apoptosis, cytotoxicity, modulation of platelet aggregation, hemorrhage, and neutrophil activation. Human neutrophils respond to LAAO by enhancing chemotaxis, and phagocytosis, and releasing reactive oxygen species (ROS) and pro-inflammatory mediators. Exosomes cellular nanovesicles play vital roles in intercellular communication, including immune responses. This study investigates the impact of Calloselasma rhodostoma snake venom-derived LAAO (Cr-LAAO) on human neutrophil exosome release, including activation patterns, exosome formation, and content. Neutrophils isolated from healthy donors were stimulated with Cr-LAAO (100 µg/mL) for 3 h, followed by exosome isolation and analysis. Results show that Cr-LAAO induces the release of exosomes with distinct protein content compared to the negative control. Proteomic analysis reveals proteins related to the regulation of immune responses and blood coagulation. This study uncovers Cr-LAAO's ability to activate human neutrophils, leading to exosome release and facilitating intercellular communication, offering insights into potential therapeutic approaches for inflammatory and immunological disorders.


Asunto(s)
Exosomas , L-Aminoácido Oxidasa , Humanos , L-Aminoácido Oxidasa/farmacología , L-Aminoácido Oxidasa/metabolismo , Neutrófilos , Exosomas/metabolismo , Peróxido de Hidrógeno/farmacología , Proteómica , Venenos de Serpiente
2.
Chem Biol Interact ; 333: 109347, 2021 Jan 05.
Artículo en Inglés | MEDLINE | ID: mdl-33259806

RESUMEN

Several reports have suggested that photobiomodulation, owing to its analgesic, anti-inflammatory, and healing effects, may be an effective therapeutic option for local effects of snakebites when the availability and accessibility of conventional serum therapy are inefficient and far from medical care centers. Although there have been studies that demonstrate the application of photobiomodulation in the treatment of local adverse events due to snakebites from snakes of the genus Bothrops, its role in the activation of leukocytes, particularly macrophages, has not been evaluated. Here, we assessed the effect of light-emitting diode (LED) treatment on macrophage activation induced by B. jararacussu venom (BjV). LED treatment caused an increase in the viability of macrophages incubated with BjV. This treatment reduced reactive oxygen species (ROS) and nitric oxide (NO) production by macrophages after incubation with BjV. However, LED treatment did not interfere with IL-1ß and IL-10 production by macrophages after incubation with BjV. In conclusion, this study showed that LED treatment has the potential to be used in combination with conventional serum therapy to prevent or minimize the progression of local to severe symptoms after Bothrops envenomation.


Asunto(s)
Bothrops , Venenos de Crotálidos/toxicidad , Terapia por Luz de Baja Intensidad/instrumentación , Macrófagos/efectos de la radiación , Semiconductores , Mordeduras de Serpientes/inmunología , Mordeduras de Serpientes/radioterapia , Animales , Supervivencia Celular/efectos de los fármacos , Interleucina-10/metabolismo , Interleucina-1beta/metabolismo , Espacio Intracelular/efectos de los fármacos , Espacio Intracelular/metabolismo , Espacio Intracelular/efectos de la radiación , Macrófagos/inmunología , Masculino , Ratones , Óxido Nítrico/biosíntesis , Especies Reactivas de Oxígeno/metabolismo , Mordeduras de Serpientes/metabolismo , Mordeduras de Serpientes/patología , Superóxidos/metabolismo
3.
Biomed Res Int ; 2014: 195356, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24724078

RESUMEN

Bothrops mattogrossensis snake is widely distributed throughout eastern South America and is responsible for snakebites in this region. This paper reports the purification and biochemical characterization of three new phospholipases A2 (PLA2s), one of which is presumably an enzymatically active Asp49 and two are very likely enzymatically inactive Lys49 PLA2 homologues. The purification was obtained after two chromatographic steps on ion exchange and reverse phase column. The 2D SDS-PAGE analysis revealed that the proteins have pI values around 10, are each made of a single chain, and have molecular masses near 13 kDa, which was confirmed by MALDI-TOF mass spectrometry. The N-terminal similarity analysis of the sequences showed that the proteins are highly homologous with other Lys49 and Asp49 PLA2s from Bothrops species. The PLA2s isolated were named BmatTX-I (Lys49 PLA2-like), BmatTX-II (Lys49 PLA2-like), and BmatTX-III (Asp49 PLA2). The PLA2s induced cytokine release from mouse neutrophils and showed cytotoxicity towards JURKAT (leukemia T) and SK-BR-3 (breast adenocarcinoma) cell lines and promastigote forms of Leishmania amazonensis. The structural and functional elucidation of snake venoms components may contribute to a better understanding of the mechanism of action of these proteins during envenomation and their potential pharmacological and therapeutic applications.


Asunto(s)
Bothrops/metabolismo , Leishmania/efectos de los fármacos , Micotoxinas/química , Micotoxinas/farmacología , Neoplasias Experimentales/tratamiento farmacológico , Venenos de Serpiente/química , Venenos de Serpiente/farmacología , Animales , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Masculino , Ratones , Micotoxinas/aislamiento & purificación , Neoplasias Experimentales/patología , Venenos de Serpiente/aislamiento & purificación , Tasa de Supervivencia , Resultado del Tratamiento
4.
Biomed Res Int ; 2013: 789689, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-23509779

RESUMEN

Bothrops asper (Squamata: Viperidae) is the most important venomous snake in Central America, being responsible for the majority of snakebite accidents. Four basic PLA2s (pMTX-I to -IV) were purified from crude venom by a single-step chromatography using a CM-Sepharose ion-exchange column (1.5 × 15 cm). Analysis of the N-terminal sequence demonstrated that pMTX-I and III belong to the catalytically active Asp49 phospholipase A2 subclass, whereas pMTX-II and IV belong to the enzymatically inactive Lys49 PLA2s-like subclass. The PLA2s isolated from Panama Bothrops asper venom (pMTX-I, II, III, and IV) are able to induce myotoxic activity, inflammatory reaction mainly leukocyte migration to the muscle, and induce J774A.1 macrophages activation to start phagocytic activity and superoxide production.


Asunto(s)
Bothrops , Macrófagos/efectos de los fármacos , Fosfolipasas A2/farmacología , Venenos de Serpiente/enzimología , Superóxidos/metabolismo , Animales , Línea Celular , Movimiento Celular , Cromatografía por Intercambio Iónico , Coagulantes/metabolismo , Edema/patología , Hemorragia/metabolismo , Inflamación , Leucocitos/citología , Macrófagos/citología , Macrófagos/metabolismo , Masculino , Ratones , Microscopía de Contraste de Fase , Panamá , Fagocitosis , Fosfolipasas A2/química
5.
J Venom Res ; 3: 28-34, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-23487552

RESUMEN

The local and systemic alterations induced by Bothrops atrox snake venom (BaV) injection in mice were studied. BaV induced superoxide production by migrated neutrophils, mast cell degranulation and phagocytosis by macrophages. Moreover, BaV caused hemorrhage in dorsum of mice after 2hr post- injection. Three hours post-injection in gastrocnemius muscle, we also observed myonecrosis, which was assessed by the determination of serum and tissue CK besides the release of urea, but not creatinine and uric acid, indicating kidney alterations. BaV also induced the release of LDH and transaminases (ALT and AST) indicating tissue and liver abnormalities. In conclusion, the data indicate that BaV induces events of local and systemic importance.

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