RESUMO
Snake envenomation is a common but neglected disease that affects millions of people around the world annually. Among venomous snake species in Brazil, the tropical rattlesnake (Crotalus durissus terrificus) accounts for the highest number of fatal envenomations and is responsible for the second highest number of bites. Snake venoms are complex secretions which, upon injection, trigger diverse physiological effects that can cause significant injury or death. The components of C. d. terrificus venom exhibit neurotoxic, myotoxic, hemotoxic, nephrotoxic, and cardiotoxic properties which present clinically as alteration of central nervous system function, motor paralysis, seizures, eyelid ptosis, ophthalmoplesia, blurred vision, coagulation disorders, rhabdomyolysis, myoglobinuria, and cardiorespiratory arrest. In this study, we focused on proteomic characterization of the cardiotoxic effects of C. d. terrificus venom in mouse models. We injected venom at half the lethal dose (LD50) into the gastrocnemius muscle. Mouse hearts were removed at set time points after venom injection (1 h, 6 h, 12 h, or 24 h) and subjected to trypsin digestion prior to high-resolution mass spectrometry. We analyzed the proteomic profiles of >1300 proteins and observed that several proteins showed noteworthy changes in their quantitative profiles, likely reflecting the toxic activity of venom components. Among the affected proteins were several associated with cellular deregulation and tissue damage. Changes in heart protein abundance offer insights into how they may work synergistically upon envenomation. SIGNIFICANCE: Venom of the tropical rattlesnake (Crotalus durissus terririficus) is known to be neurotoxic, myotoxic, nephrotoxic and cardiotoxic. Although there are several studies describing the biochemical effects of this venom, no work has yet described its proteomic effects in the cardiac tissue of mice. In this work, we describe the changes in several mouse cardiac proteins upon venom treatment. Our data shed new light on the clinical outcome of the envenomation by C. d. terrificus, as well as candidate proteins that could be investigated in efforts to improve current treatment approaches or in the development of novel therapeutic interventions in order to reduce mortality and morbidity resulting from envenomation.
Assuntos
Venenos de Crotalídeos , Síndromes Neurotóxicas , Mordeduras de Serpentes , Animais , Venenos de Crotalídeos/química , Crotalus/metabolismo , Humanos , Camundongos , Proteínas/metabolismo , Proteômica , Mordeduras de Serpentes/terapiaRESUMO
Snake envenomation is a common but neglected disease that affects millions of people around the world annually. Among venomous snake species in Brazil, the tropical rattlesnake (Crotalus durissus terrificus) accounts for the highest number of fatal envenomations and is responsible for the second highest number of bites. Snake venoms are complex secretions which, upon injection, trigger diverse physiological effects that can cause significant injury or death. The components of C. d. terrificus venom exhibit neurotoxic, myotoxic, hemotoxic, nephrotoxic, and cardiotoxic properties which present clinically as alteration of central nervous system function, motor paralysis, seizures, eyelid ptosis, ophthalmoplesia, blurred vision, coagulation disorders, rhabdomyolysis, myoglobinuria, and cardiorespiratory arrest. In this study, we focused on proteomic characterization of the cardiotoxic effects of C. d. terrificus venom in mouse models. We injected venom at half the lethal dose (LD50) into the gastrocnemius muscle. Mouse hearts were removed at set time points after venom injection (1 h, 6 h, 12 h, or 24 h) and subjected to trypsin digestion prior to high-resolution mass spectrometry. We analyzed the proteomic profiles of >1300 proteins and observed that several proteins showed noteworthy changes in their quantitative profiles, likely reflecting the toxic activity of venom components. Among the affected proteins were several associated with cellular deregulation and tissue damage. Changes in heart protein abundance offer insights into how they may work synergistically upon envenomation. Significance Venom of the tropical rattlesnake (Crotalus durissus terririficus) is known to be neurotoxic, myotoxic, nephrotoxic and cardiotoxic. Although there are several studies describing the biochemical effects of this venom, no work has yet described its proteomic effects in the cardiac tissue of mice. In this work, we describe the changes in several mouse cardiac proteins upon venom treatment. Our data shed new light on the clinical outcome of the envenomation by C. d. terrificus, as well as candidate proteins that could be investigated in efforts to improve current treatment approaches or in the development of novel therapeutic interventions in order to reduce mortality and morbidity resulting from envenomation.
RESUMO
As técnicas de cultura celular são utilizadas desde o começo do século XX e vêm ganhando espaço em diferentes áreas de pesquisa. Um exemplo é na farmacodinâmica, em estudos cujo objetivo é observar o efeito de drogas, biofarmacos e compostos tóxicos, como venenos de serpentes. O veneno de serpentes é composto por uma mistura complexa de proteínas, peptídeos, aminoácidos, nucleotídeos, lipídeos e carboidratos que apresentam uma gama de ações isoladas ou em conjunto. Diversos estudos têm sido realizados a fim de analisar o potencial uso de venenos para o tratamento de doenças tais como o câncer utilizando diferentes metodologias, sendo duas delas: a cultura celular utilizando linhagens celulares tumorais e a proteômica. Proteômica consiste na análise dos conjuntos de proteínas e suas isoformas expressas em uma amostra biológica, podendo ser um organismo, tecido ou célula. A análise proteômica baseada em espectrometria de massas é uma técnica que separa e identifica proteínas por meio da medição de sua massa, medida com base na relação massa/carga de íons em fase gasosa. O presente trabalho busca analisar e caracterizar o perfil morfológico e proteômico das linhagens celulares tumorais sob efeito do veneno da serpente Bothrops jararaca a partir da cultura de células tumorais MCF-7 e MDA-MB-231. Foram observadas diferentes morfologias entre as diferentes linhagens celulares após tratamento com baixa (0,63 mg/mL) e alta dose sub-citotóxica (2,5 mg/mL) de veneno. A análise por espectrometria de massas permitiu a identificação de diferentes proteínas em cada uma das amostras e a presença de proteínas comuns entre amostras controle e tratadas com as diferentes doses de veneno. Trabalhos subsequentes utilizando doses diferentes de veneno e tempo de tratamento permitirão melhor caracterizar o perfil proteômico das diferentes linhagens tumorais sob efeito do veneno de B. jararaca.