Avaliação dos efeitos da Relaxina exógena sobre olho seco em modelo de síndrome de Sjögren (SjS)
Data
2022-12-12
Autores
Carvalho, Lucimeire Nova de 
Orientadores
Gomes, José Álvaro Pereira
Tipo
Tese de doutorado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
O objetivo deste trabalho foi avaliar o potencial da relaxina (RLX) no tratamento da doença do olho seco (DOS) em modelo experimental animal com SjS. Métodos Camundongos NOD diagnosticados com SjS foram tratados durante trinta dias, 4x/dia, com colírio a base de RLX (100ng/ml). Os animais foram submetidos à análise clínica para verificar a produção lacrimal e a ceratite antes, durante e após o tratamento. Tecidos oculares, glândulas de Meibomius, glândula lacrimal exorbitária e glândula de Harder foram retirados e submetidos a diferentes técnicas. As glândulas lacrimais (GL) foram analisadas por PCR array para verificar as citocinas inflamatórias IL-1b, IL-6, IL-10, IL-12a, IL-17a, TNFα e INFϒ e também RLX e RXFP1. A técnica da imunofluorescência foi usada para detecção do hormônio RLX e seu receptor RXFP1 nos órgãos citados e análise histológica foi empregada para verificar possíveis diferenças entre os órgãos citados anteriormente. Resultados: O grupo tratado apresentou melhora significativa em todos os aspectos clínicos analisados, como aumento da produção lacrimal, reversão da ceratite, inclusive no olho que não recebeu o tratamento (contralateral). Todos os tecidos analisados mostraram marcação positiva para a RLX e seu receptor (RXFP1). Quando verificado o aumento na expressão da RLX e RXFP1, o grupo tratado com RLX mostrou um aumento significante na expressão gênica em relação aos demais grupos. Ainda por RT-PCR, as análises das citocinas IL1b, IL-6, IL-12, IL-17 e TNF estavam significantemente menores no grupo tratado em relação aos demais. Conclusão: Esses resultados sugerem que a RLX exógena apresenta potencial ação no tratamento da DOS, podendo ser usada em forma de colírio, uma vez que houve melhora no quadro clínico e diminuição das citocinas inflamatórias típicas da DOS por SjS.
The objective of this work was to evaluate the potential of relaxin (RLX) in the treatment of dry eye disease (DED) in an experimental animal model of SjS. Methods NOD mice diagnosed with SjS were treated for thirty days, 4x/day, with RLX-based eye drops (100ng/ml). The animals were submitted to clinical analysis to verify tear production and keratitis before, during and after treatment. Ocular tissues, meibomian glands, exorbital lacrimal gland and Harder's gland were removed and subjected to different techniques. Lacrimal glands were analyzed by PCR array to verify the inflammatory cytokines IL-1b, IL-6, IL-10, IL-12a, IL-17a, TNFα and INFϒ and also RLX and RXFP1. The immunofluorescence technique was used to detect the RLX hormone and its RXFP1 receptor in organs and histological analysis was used to verify possible differences between the aforementioned organs. Results: The treated group showed significant improvement in all clinical aspects analyzed, such as increased tear production, reversal of keratitis, including in the eye that did not receive the contralateral treatment. All analyzed tissues showed positive staining for RLX and its receptor (RXFP1). When the increase in RLX and RXFP1 expression was verified, the RLX-treated group showed a significant increase in gene expression in relation to the other groups. As for RT-PCR, the analysis of cytokines IL1b, IL-6, IL-12, IL- 17, and TNF were significantly lower in the treated group than in the others. Conclusion: These results suggest that exogenous RLX has potential in the treatment of DED and can be used in the form of eye drops, since there was an improvement in the clinical picture and decreased the typical inflammatory cytokines of DES by SjS.
The objective of this work was to evaluate the potential of relaxin (RLX) in the treatment of dry eye disease (DED) in an experimental animal model of SjS. Methods NOD mice diagnosed with SjS were treated for thirty days, 4x/day, with RLX-based eye drops (100ng/ml). The animals were submitted to clinical analysis to verify tear production and keratitis before, during and after treatment. Ocular tissues, meibomian glands, exorbital lacrimal gland and Harder's gland were removed and subjected to different techniques. Lacrimal glands were analyzed by PCR array to verify the inflammatory cytokines IL-1b, IL-6, IL-10, IL-12a, IL-17a, TNFα and INFϒ and also RLX and RXFP1. The immunofluorescence technique was used to detect the RLX hormone and its RXFP1 receptor in organs and histological analysis was used to verify possible differences between the aforementioned organs. Results: The treated group showed significant improvement in all clinical aspects analyzed, such as increased tear production, reversal of keratitis, including in the eye that did not receive the contralateral treatment. All analyzed tissues showed positive staining for RLX and its receptor (RXFP1). When the increase in RLX and RXFP1 expression was verified, the RLX-treated group showed a significant increase in gene expression in relation to the other groups. As for RT-PCR, the analysis of cytokines IL1b, IL-6, IL-12, IL- 17, and TNF were significantly lower in the treated group than in the others. Conclusion: These results suggest that exogenous RLX has potential in the treatment of DED and can be used in the form of eye drops, since there was an improvement in the clinical picture and decreased the typical inflammatory cytokines of DES by SjS.
Descrição
Citação
CARVALHO, Lucimeire Nova de. Avaliação dos efeitos da Relaxina exógena sobre olho seco em modelo de síndrome de Sjögren (SjS). São Paulo, 2022. 95 p. Tese (Doutorado em Oftalmologia e Ciências Visuais) – Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, 2022.