Proteína anti-inflamatória Anexina A1 como alternativa terapêutica na uveíte autoimune experimental
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Data
2022-03-03
Autores
Costa, Sara de Souza
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
A uveíte autoimune é uma importante inflamação ocular relacionada à perda visual. A proteína Anexina A1 (AnxA1) e seu peptídeo mimético, Ac2-26, foram pesquisados em diferentes condições oculares com perspectivas clínicas. Assim, investigamos, in vivo, os efeitos do Ac2-26 para prevenção e protocolos terapêuticos na uveíte autoimune experimental (EAU). Para induzir EAU, a proteína de ligação a retinoides interfotorreceptores bovina (IRBP) em adjuvante completo de Freund foi inoculada na pata direita de ratos Lewis. Adicionalmente, a toxina de Bordetella pertussis foi injetada intraperitonealmente (i.p.). Ac2-26 foi administrado diariamente (i.p.) seguindo os protocolos preventivos (a partir do dia 1) ou terapêuticos (a partir do dia 8). Ambos os tratamentos foram realizados até o dia 13 após a indução da EAU e a gravidade da EAU foi avaliada diariamente. A quantificação de leucócitos foi realizada em humor aquoso (AqH). As expressões de AnxA1, receptor de formil-peptídeo-2 (fpr2), ciclooxigenase-2 (COX-2) e quantificação de macrófagos (ED1) foram avaliados por imuno-histoquímica. Linfócitos CD4 e CD8, AnxA1 e fpr2 foram estudados por Western blotting. Interferon (IFN)-γ, fator de necrose tumoral (TNF)-α, interleucinas (IL-2, IL-4, IL-10, IL-17) e proteína quimioatraente de monócitos (MCP)-1 foram medidos em sobrenadante de macerado ocular. Clinicamente, os olhos dos ratos induzidos por EAU apresentaram características macroscópicas alteradas e, nos grupos tratados, os parâmetros normais foram restabelecidos. Leucócitos, predominantemente linfócitos, foram encontrados no grupo EAU não tratado, enquanto a redução de CD4, CD8 e ED1 ocorreu nos animais tratados. Além disso, Ac2-26 diminuiu a imunorreatividade de AnxA1, fpr2 e COX-2 em segmentos oculares e sobrenadantes. Além disso, níveis reduzidos de IL-2, IFN-γ e IL-10 foram detectados em grupos tratados com Ac2-26. Entre os tratamentos, a administração terapêutica foi mais eficaz. Os dados mostram os efeitos benéficos do Ac2-26 em EAU com potencial para aplicação terapêutica.
Autoimmune uveitis is an important ocular inflammation related to visual loss. Annexin A1 (AnxA1) protein and its mimetic peptide, Ac2-26, have been researched in different eye conditions with clinical perspectives. Thus, we investigated, in vivo, the Ac2-26 effects for prevention and therapeutic protocols in the experimental autoimmune uveitis (EAU). To induce EAU, bovine interphotoreceptor retinoid-binding protein (IRBP) in complete Freund's adjuvant was inoculated in the right paw of Lewis rats. Additionally, Bordetella pertussis toxin was injected intraperitoneally (i.p.). Ac2-26 was daily administered (i.p.) following the preventive (from day 1) or therapeutic (from day 8) protocols. Both treatments were performed until day 13 after EAU induction and EAU severity was daily assessed. The quantification of leukocytes was performed in aqueous humor (AqH). AnxA1, formyl-peptide receptor-2 (fpr2), cyclooxygenase-2 (COX 2) expressions and macrophage quantification (ED1) were evaluated by immunohistochemistry. CD4 and CD8 lymphocytes, AnxA1 and fpr2 were studied by Western blotting. Interferon (IFN)-γ, tumor necrosis factor (TNF)-α, interleukins (IL-2, IL-4, IL-10, IL-17) and monocyte chemoattractant protein (MCP)-1 were measured in ocular macerate supernatant. Clinically, the eyes of the EAU-induced rats showed altered macroscopic characteristics, and in the treated groups, normal parameters were reestablished. Leukocytes, predominantly lymphocytes, were found in the untreated-EAU group, while reduction of CD4, CD8 and ED1 occurred in treated animals. In addition, Ac2-26 decreased AnxA1, fpr2 and COX-2 immunoreactivity in ocular segments and supernatants. Furthermore, reduced levels of IL-2, IFN-γ and IL-10 were detected in Ac2-26-treated groups. Between the treatments, therapeutic administration was more effective. Data show the beneficial effects of Ac2-26 in EAU with potential for therapeutic application.
Autoimmune uveitis is an important ocular inflammation related to visual loss. Annexin A1 (AnxA1) protein and its mimetic peptide, Ac2-26, have been researched in different eye conditions with clinical perspectives. Thus, we investigated, in vivo, the Ac2-26 effects for prevention and therapeutic protocols in the experimental autoimmune uveitis (EAU). To induce EAU, bovine interphotoreceptor retinoid-binding protein (IRBP) in complete Freund's adjuvant was inoculated in the right paw of Lewis rats. Additionally, Bordetella pertussis toxin was injected intraperitoneally (i.p.). Ac2-26 was daily administered (i.p.) following the preventive (from day 1) or therapeutic (from day 8) protocols. Both treatments were performed until day 13 after EAU induction and EAU severity was daily assessed. The quantification of leukocytes was performed in aqueous humor (AqH). AnxA1, formyl-peptide receptor-2 (fpr2), cyclooxygenase-2 (COX 2) expressions and macrophage quantification (ED1) were evaluated by immunohistochemistry. CD4 and CD8 lymphocytes, AnxA1 and fpr2 were studied by Western blotting. Interferon (IFN)-γ, tumor necrosis factor (TNF)-α, interleukins (IL-2, IL-4, IL-10, IL-17) and monocyte chemoattractant protein (MCP)-1 were measured in ocular macerate supernatant. Clinically, the eyes of the EAU-induced rats showed altered macroscopic characteristics, and in the treated groups, normal parameters were reestablished. Leukocytes, predominantly lymphocytes, were found in the untreated-EAU group, while reduction of CD4, CD8 and ED1 occurred in treated animals. In addition, Ac2-26 decreased AnxA1, fpr2 and COX-2 immunoreactivity in ocular segments and supernatants. Furthermore, reduced levels of IL-2, IFN-γ and IL-10 were detected in Ac2-26-treated groups. Between the treatments, therapeutic administration was more effective. Data show the beneficial effects of Ac2-26 in EAU with potential for therapeutic application.
Descrição
Palavras-chave
Inflamação ocular, Uveíte autoimune, Anexina A1, Mediadores inflamatórios, Eye inflammation, Autoimmune uveitis, Annexin A1, Inflammatory mediators