RESUMO
Despite years of effort, current therapies for diabetic wounds are still not fully efficacious. Emerging evidence has suggested that microRNAs (miRNAs) play key roles in multiple physiological and pathological processes in eukaryotes, and could potentially be powerful therapeutic tools. This study investigated the differential expression profiling of miRNAs in cutaneous wounds in streptozotocin-induced diabetic rats and normal rats, and its significance in diabetic wound healing. Using microarrays, 18 miRNAs were identified as being upregulated and 65 as being downregulated in the diabetic group. The miRNA profiling results were validated by quantitative reverse transcriptase polymerase chain reaction. Finally, functional annotation analysis using the DAVID and miR2Subpath databases revealed that the differentially expressed miRNAs were involved in MAPK signaling pathways, the Wnt signaling pathway, and other signaling pathways that may be closely linked to wound healing. This study provides an experimental foundation for further investigation of mechanisms that underlie poor diabetic wound healing, and of miRNA-based therapies that are associated with wound healing.
Assuntos
Complicações do Diabetes , Diabetes Mellitus Experimental , MicroRNAs/genética , Transcriptoma , Ferimentos e Lesões/etiologia , Animais , Biologia Computacional , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Masculino , Anotação de Sequência Molecular , Fenótipo , Ratos , Reprodutibilidade dos Testes , Transdução de Sinais , Úlcera Cutânea/etiologia , Úlcera Cutânea/genética , Úlcera Cutânea/metabolismo , Cicatrização/genética , Ferimentos e Lesões/genética , Ferimentos e Lesões/metabolismoRESUMO
The host immune response has a critical role not only in protection from human leishmaniasis but also in promoting disease severity. Although candidate gene approaches in mouse models of leishmaniasis have been extremely informative, a global understanding of the immune pathways active in lesions from human patients is lacking. To address this issue, genome-wide transcriptional profiling of Leishmania braziliensis-infected cutaneous lesions and normal skin controls was carried out. A signature of the L. braziliensis skin lesion was defined, which includes over 2,000 differentially regulated genes. Pathway-level analysis of this transcriptional response revealed key biological pathways present in cutaneous lesions, generating a testable 'metapathway' model of immunopathology and providing new insights for treatment of human leishmaniasis.
Assuntos
Genômica/métodos , Leishmania braziliensis/imunologia , Leishmaniose Cutânea , Úlcera Cutânea , Adolescente , Adulto , Feminino , Humanos , Inflamassomos/genética , Inflamassomos/imunologia , Leishmaniose Cutânea/genética , Leishmaniose Cutânea/imunologia , Leishmaniose Cutânea/parasitologia , Masculino , Pessoa de Meia-Idade , Psoríase/genética , Psoríase/imunologia , Pele/imunologia , Pele/parasitologia , Úlcera Cutânea/genética , Úlcera Cutânea/imunologia , Úlcera Cutânea/parasitologia , Transcrição Gênica/imunologia , Transcriptoma/imunologia , Adulto JovemRESUMO
Classic and molecular (polymerase chain reaction--PCR) techniques were used to diagnose American cutaneous leishmaniasis in 149 dogs from an area in the northwest of Paraná State, Brazil, where an American cutaneous leishmaniasis outbreak occurred in 2002. The results were compared to a set of previously obtained results. Twenty-five dogs had positive indirect immunofluorescence (IIF) (titers > or = 40), including two animals with suggestive lesions. The percentage of dogs with positive IIF was similar to that found in a previous study. The cultures of the lesion, blood and bone marrow were negative for Leishmania. A direct search for the parasite in the lesions proved negative, although PCR tests were positive. The PCR did not detect the DNA of Leishmania (Viannia) in the blood, even for those that had positive PCR in a previous study. The follow up of the 27 dogs showed that the majority of them had maintained the same levels of antibodies that had been detected previously. There was a reduction in the number of dogs with lesions, probably due to the transmission control measures that were adopted after the outbreak.
Assuntos
Reservatórios de Doenças/veterinária , Doenças do Cão/diagnóstico , Leishmania braziliensis , Leishmaniose Cutânea/veterinária , Animais , Anticorpos Antiprotozoários/sangue , Anticorpos Antiprotozoários/genética , Medula Óssea/parasitologia , Medula Óssea/patologia , Brasil/epidemiologia , Meios de Cultura , DNA de Protozoário/sangue , DNA de Protozoário/genética , Surtos de Doenças/veterinária , Reservatórios de Doenças/parasitologia , Reservatórios de Doenças/estatística & dados numéricos , Doenças do Cão/epidemiologia , Doenças do Cão/parasitologia , Cães , Feminino , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Leishmania braziliensis/genética , Leishmania braziliensis/imunologia , Leishmania braziliensis/isolamento & purificação , Leishmaniose Cutânea/sangue , Leishmaniose Cutânea/epidemiologia , Leishmaniose Cutânea/genética , Masculino , Reação em Cadeia da Polimerase/veterinária , População Rural , Úlcera Cutânea/genética , Úlcera Cutânea/patologia , Úlcera Cutânea/veterinária , Fatores de TempoRESUMO
Classic and molecular (polymerase chain reaction - PCR) techniques were used to diagnose American cutaneous leishmaniasis in 149 dogs from an area in the northwest of Paraná State, Brazil, where an American cutaneous leishmaniasis outbreak occurred in 2002. The results were compared to a set of previously obtained results. Twenty-five dogs had positive indirect immunofluorescence (IIF) (titers > 40), including two animals with suggestive lesions. The percentage of dogs with positive IIF was similar to that found in a previous study. The cultures of the lesion, blood and bone marrow were negative for Leishmania. A direct search for the parasite in the lesions proved negative, although PCR tests were positive. The PCR did not detect the DNA of Leishmania (Viannia) in the blood, even for those that had positive PCR in a previous study. The follow up of the 27 dogs showed that the majority of them had maintained the same levels of antibodies that had been detected previously. There was a reduction in the number of dogs with lesions, probably due to the transmission control measures that were adopted after the outbreak.
Neste estudo, utilizaram-se técnicas clássicas e moleculares (reação em cadeia da polimerase - PCR) para o diagnóstico da leishmaniose tegumentar americana em 149 cães de uma área no noroeste do Estado do Paraná, Brasil, onde ocorreu um surto de leishmaniose tegumentar americana em 2002; os resultados foram comparados aos obtidos anteriormente. Vinte e cinco cães tiveram a imunofluorescência indireta (IFI) positiva (títulos > 40), incluindo dois animais com lesão sugestiva. O percentual de cães com IFI positiva foi semelhante aos encontrados nos inquéritos anteriores. As culturas dos materiais de lesão, sangue e medula óssea foram negativas para Leishmania. A pesquisa direta do parasito em lesão foi negativa, no entanto a PCR foi positiva. A PCR não detectou DNA de Leishmania (Viannia) no sangue dos cães estudados, mesmo naqueles que tiveram PCR positiva no estudo anterior. O acompanhamento de 27 animais mostrou que a maioria deles permaneceu com os mesmos níveis de anticorpos detectados anteriormente. Houve redução do número de cães com lesões, provavelmente em virtude das medidas de controle da transmissão adotadas após o surto de 2002.