Avaliação morfofisiológica e imunológica in vitro dos efeitos dos extratos salivares de carrapatos Rhipicephalus sanguineus sensu lato (Acari: Ixodidae)
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Data
2017-08-31
Autores
Abreu, Marina Rodrigues de [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
A espécie Rhipicephalus sanguineus s. l., mais conhecida como “carrapato do cão”, possui uma grande importância médico-veterinária, não só por espoliar seus hospedeiros, mas também por ser um potencial vetor de agentes patogênicos. O sucesso biológico desses ectoparasitas deve-se à produção de uma saliva complexa produzida pelas glândulas salivares, cujos compostos são capazes de modular o sistema imune inflamatório e hemostático dos hospedeiros. No entanto, mesmo com o conhecimento adquirido sobre o potencial da saliva no sistema imune-inflamatório dos hospedeiros, pouco se estudou sobre os mecanismos de ação dos extratos glandulares nessa resposta, e as alterações morfofisiológicas envolvidas. Desta forma, o presente projeto propôs a análise dos efeitos in vitro de extratos de glândulas salivares de fêmeas de carrapatos R. sanguineus s. l. com 2 e 4 dias de alimentação (EG2 e EG4) sobre macrófagos peritoneais de camundongos BALB/c e células da linhagem J774, observando possíveis variações na atividade e na morfofisiologia destas células em função das diferentes exposições ao extrato. Os resultados demonstraram que apenas a concentração de 4 µg/mL do extrato EG2 apresentou citotoxicidade sobre células da linhagem J774 quando expostas por 48 horas; as concentrações de 4 e 2 µg/mL de ambos os extratos estimularam perfis pró inflamatórios em macrófagos peritoneais de camundongos BALB/c; a concentração de 2 µg/mL no tempo de 48 horas de exposição e 1 µg/mL no de 24 horas de exposição do EG2 e as concentrações 2 e 1 µg/mL o EG4 em 48 horas de exposição também estimularam atividade pró-inflamatória de células da linhagem J774; as demais concentrações dos extratos EG2 e EG4 tiveram atividade imunomodulatória sobre ambas linhagens, como demonstrado pela diminuição de liberação de NO, concomitante com a diminuição da secreção de citocinas pró-inflamatórias (IL2, IL-6 e TNF-α), aumento da síntese de IL-10, e confirmada pela análise morfológica sob microscopia de luz e varredura.
The species Rhipicephalus sanguineus s. l., better known as "dog tick", has a great medical-veterinary importance, not only for spolling its hosts, but also because it is a potential vector of pathogens. Biological success of these ectoparasites is due to the production of a complex saliva produced by salivary glands, whose compounds are able to modulate the immune system inflammatory and hemostatic properties of the hosts. However, even with the knowledge acquired on the potential of saliva in the immune-inflammatory system of the hosts, little was studied about the mechanisms of action of glandular extracts in this response, and the morphophysiological changes involved. In this way, the present project proposed analysis of the in vitro effects of extracts of salivary glands from female ticks R. sanguineus s. l. with 2 and 4 days of feeding (EG2 and EG4) on peritoneal macrophages of BALB / c mice and J774 cells, noting possible variations in the activity and in the morphophysiology of these cells due to the different exposures to extract. The results demonstrated that only the concentration of 4 μg / mL of the extract EG2 showed cytotoxicity on cells of the J774 lineage when exposed by 48 hours; the concentrations of 4 and 2 μg / mL of both extracts stimulated pro-inflammatory response in peritoneal macrophages of BALB / c mice; the concentration of 2 μg / mL at the 48 hour exposure time and 1 μg / mL at the 24 hour exposure time of the EG2 and concentrations 2 and 1 μg / mL EG4 at 48 hours of exposure also stimulated pro-inflammatory activity of cells of the J774 lineage; the rest concentrations of extracts EG2 and EG4 had immunomodulatory activity on both lines, as demonstrated by the decrease in NO release, concomitant with decreased secretion of pro-inflammatory cytokines (IL2, IL-6 and TNF-α), increased IL-10 synthesis, and confirmed by morphological analysis under light microscopy and scanning.
The species Rhipicephalus sanguineus s. l., better known as "dog tick", has a great medical-veterinary importance, not only for spolling its hosts, but also because it is a potential vector of pathogens. Biological success of these ectoparasites is due to the production of a complex saliva produced by salivary glands, whose compounds are able to modulate the immune system inflammatory and hemostatic properties of the hosts. However, even with the knowledge acquired on the potential of saliva in the immune-inflammatory system of the hosts, little was studied about the mechanisms of action of glandular extracts in this response, and the morphophysiological changes involved. In this way, the present project proposed analysis of the in vitro effects of extracts of salivary glands from female ticks R. sanguineus s. l. with 2 and 4 days of feeding (EG2 and EG4) on peritoneal macrophages of BALB / c mice and J774 cells, noting possible variations in the activity and in the morphophysiology of these cells due to the different exposures to extract. The results demonstrated that only the concentration of 4 μg / mL of the extract EG2 showed cytotoxicity on cells of the J774 lineage when exposed by 48 hours; the concentrations of 4 and 2 μg / mL of both extracts stimulated pro-inflammatory response in peritoneal macrophages of BALB / c mice; the concentration of 2 μg / mL at the 48 hour exposure time and 1 μg / mL at the 24 hour exposure time of the EG2 and concentrations 2 and 1 μg / mL EG4 at 48 hours of exposure also stimulated pro-inflammatory activity of cells of the J774 lineage; the rest concentrations of extracts EG2 and EG4 had immunomodulatory activity on both lines, as demonstrated by the decrease in NO release, concomitant with decreased secretion of pro-inflammatory cytokines (IL2, IL-6 and TNF-α), increased IL-10 synthesis, and confirmed by morphological analysis under light microscopy and scanning.
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Palavras-chave
Ácaro, Morfologia, Imunologia, Glândulas salivares, Macrófagos, Imunomodulação