Avaliação de terapias antifibróticas associadas aos antifúngicos itraconazol e cotrimoxazol em modelo murino de paracoccidioidomicose pulmonar
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Data
2017-08-01
Autores
Finato, Angela Carolina [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Paracoccidioidomicose (PCM) é uma micose sistêmica causada por fungos do gênero Paracoccidioides; suas principais formas clínicas são aguda/subaguda, crônica e residual. A PCM é uma doença restrita a países da América Latina com maior incidência no Brasil, especialmente entre os trabalhadores rurais. A maioria dos pacientes com a forma crônica da doença, mesmo após tratamento eficaz, apresentam sequelas, incluindo fibrose pulmonar e adrenal. Os problemas sociais, econômicos e psicológicos desencadeados pela fibrose pulmonar são subestimados; além disso, a fibrose na PCM permanece negligenciada, uma vez que não há tratamento. Dessa forma, o estudo teve por objetivo investigar a influência de drogas com potencial antifibrótico (pentoxifilina - PTX, azitromicina - AZT e talidomida - Thal) associadas aos tratamentos antifúngicos com itraconazol - ITC e cotrimoxazol - CMX em modelo murino de PCM pulmonar. Para tanto, camundongos BALB/c machos foram inoculados com leveduras do isolado 326 de P. brasiliensis e após 8 semanas de infecção foi dado início aos esquemas terapêuticos: PTX/ITC, PTX/CMX, AZT/ITC, AZT/CMX, Thal/ITC e Thal/CMX. Após 8 semanas de tratamento, os animais foram eutanasiados a fim de se avaliar a deposição de fibras colágenas, produção de hidroxiprolina, recuperação de fungos viáveis e a porcentagem das áreas com lesão nos pulmões e peso corporal. Visando identificar os mecanismos envolvidos foi avaliada a produção de TGF-β1, CCL3, IFN-γ, TNF-α, IL-10, VEGF, IL-6, IL-1β, IL-17 e IL-2 no homogenato dos pulmões. Nossos achados revelaram que os camundongos infectados tratados com PTX/ITC mostraram redução nos níveis de hidroxiprolina associada a menor produção de IL-6, IL-17 e TGF-β1 e níveis mais elevados de IL-10 em comparação ao tratamento apenas com ITC. De forma semelhante, os camundongos infectados tratados com AZT/CMX apresentaram menos fibrose pulmonar associada a níveis reduzidos de TGF-β1 e aumento de TNF-α e IL-10 do que os camundongos infectados tratados com CMX. Ambos os grupos em que os camundongos foram tratados com ITC/Thal e CMX/Thal mostraram perda de peso intensa, aumento da deposição de fibras reticulares, altos níveis de CCL3, IFN-γ e VEGF e níveis diminuídos de IL-6, IL-1β, IL-17 e TGF-β1 em relação aos camundongos infectados tratados com ITC ou CMX. Dessa forma, nossos achados reforçam o papel antifibrótico da PTX na PCM pulmonar quando associado ao ITC. Além disso, nossos resultados apontaram a AZT como candidata a droga antifibrótica em associação com CMX e a ineficácia da Thal, no tratamento da PCM pulmonar.
Paracoccidioidomycosis (PCM) is a systemic mycosis caused by fungi of Paracoccidioides genus; the main clinical forms are acute/subacute, chronic and residual. PCM is a disease restricted to Latin American countries with a higher incidence in Brazil, especially among rural workers. Most patients with the chronic form, even after effective treatment, present sequelae, including pulmonary and adrenal fibrosis. The social, economic and psychological problems triggered by pulmonary sequels are underestimated. In addition, fibrosis in PCM remains neglected, since there is no treatment. The aim of this study was to investigate the influence of antifibrotic drugs (pentoxifylline - PTX, azithromycin - AZT and thalidomide - Thal) associated with antifungal treatments with itraconazole - ITC and cotrimoxazole - CMX in a murine model of pulmonary PCM. Male BALB/c mice were inoculated with P. brasiliensis “isolated 326” and after 8 weeks of infection the treatment were started: PTX/ITC, PTX/CMX, AZT/ITC, AZT/CMX, Thal/ITC and Thal/CMX. After 8 weeks of treatment, the mice were euthanized in order to evaluate the deposition of collagen fibers, hydroxyproline production, recovery of viable fungi and the percentage of areas with injury in lung and body weight. In order to identify the mechanisms involved, the production of TGF-β1, CCL3, IFN-γ, TNF-α, IL-10, VEGF, IL-6, IL-1β, IL-17 and IL-2 in the lung homogenate was evaluated. Our findings revealed that infected mice treated with PTX/ITC showed a reduction in hydroxyproline levels associated with lower production of IL-6, IL-17 and TGF-β1 and higher levels of IL-10 compared to treatment with ITC. Similarly, infected mice treated with AZT/CMX had less lung fibrosis associated with reduced levels of TGF-β1 and increased TNF-α and IL-10 than infected mice treated with CMX. Both groups in which the mice were treated with Thal/ITC and Tha/CMX showed intense weight loss, increased deposition of reticulin fiber, high levels of CCL3, IFN-γ and VEGF and decreased levels of IL-6, IL- 1β, IL-17 and TGF-β1 than to the infected mice treated with ITC or CMX. Thus, our findings reinforce the antifibrotic role of PTX in pulmonary PCM when associated with ITC. Besides, our results point out the AZT as a candidate for antifibrotic drug in association with CMX and the inefficacy of Thal in the treatment of pulmonary PCM.
Paracoccidioidomycosis (PCM) is a systemic mycosis caused by fungi of Paracoccidioides genus; the main clinical forms are acute/subacute, chronic and residual. PCM is a disease restricted to Latin American countries with a higher incidence in Brazil, especially among rural workers. Most patients with the chronic form, even after effective treatment, present sequelae, including pulmonary and adrenal fibrosis. The social, economic and psychological problems triggered by pulmonary sequels are underestimated. In addition, fibrosis in PCM remains neglected, since there is no treatment. The aim of this study was to investigate the influence of antifibrotic drugs (pentoxifylline - PTX, azithromycin - AZT and thalidomide - Thal) associated with antifungal treatments with itraconazole - ITC and cotrimoxazole - CMX in a murine model of pulmonary PCM. Male BALB/c mice were inoculated with P. brasiliensis “isolated 326” and after 8 weeks of infection the treatment were started: PTX/ITC, PTX/CMX, AZT/ITC, AZT/CMX, Thal/ITC and Thal/CMX. After 8 weeks of treatment, the mice were euthanized in order to evaluate the deposition of collagen fibers, hydroxyproline production, recovery of viable fungi and the percentage of areas with injury in lung and body weight. In order to identify the mechanisms involved, the production of TGF-β1, CCL3, IFN-γ, TNF-α, IL-10, VEGF, IL-6, IL-1β, IL-17 and IL-2 in the lung homogenate was evaluated. Our findings revealed that infected mice treated with PTX/ITC showed a reduction in hydroxyproline levels associated with lower production of IL-6, IL-17 and TGF-β1 and higher levels of IL-10 compared to treatment with ITC. Similarly, infected mice treated with AZT/CMX had less lung fibrosis associated with reduced levels of TGF-β1 and increased TNF-α and IL-10 than infected mice treated with CMX. Both groups in which the mice were treated with Thal/ITC and Tha/CMX showed intense weight loss, increased deposition of reticulin fiber, high levels of CCL3, IFN-γ and VEGF and decreased levels of IL-6, IL- 1β, IL-17 and TGF-β1 than to the infected mice treated with ITC or CMX. Thus, our findings reinforce the antifibrotic role of PTX in pulmonary PCM when associated with ITC. Besides, our results point out the AZT as a candidate for antifibrotic drug in association with CMX and the inefficacy of Thal in the treatment of pulmonary PCM.
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Palavras-chave
Antifibróticos, Azitromicina, Fibrose pulmonar, Pentoxifilina, Talidomida, Antifibrotic, Azithromycin, Pentoxifylline, Pulmonary fibrosis, Thalidomide