Avaliação da expressão de miRNAs circulantes e da articulação tibiotársica em equinos
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Data
2022-02-17
Autores
Denadai, Daniela Scantamburlo [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
A osteoartrite é uma das maiores causas de morbidade em equinos. É caracterizada pela degeneração e proliferação óssea, lesões nas cartilagens e espessamento da membrana sinovial e da cápsula articular. O estudo de miRNAs é um campo emergente na determinação dos mecanismos que regulam enfermidades inflamatórias, podendo-se criar modelos para estudar as vias de sinalização para enfermidades articulares. Foram utilizados seis equinos atletas da raça Quarto de Milha, divididos em dois grupos: Grupo Controle (CT) - sem qualquer enfermidade ósteo-articular, e Grupo Osteoartrite (OA) - diagnosticados com osteoartrite tibiotársica primária traumática subclínica. Foram coletadas amostras de sangue venoso e de líquido sinovial para realizar a extração do RNAtotal por meio do kit “Alternative Protocol for Extraction of RNA from Cells Captured on LeukoLOCK Filters Using TRI Reagent®” (Thermo Fisher Scientific®). O microarranjo foi realizado com o kit “Affymetrix® miRNA 4.1 Array Strip” e equipamento “GeneAtlas Affymetrix®”. As análises estatísticas foram realizadas com o programa “Transcriptome Analysis Console (TAC)”. Quando comparados os grupos, foram identificados 42 miRNAs diferencialmente expressos no sangue dos equinos (36 regulados positivamente e 6 regulados negativamente). Já no líquido sinovial, foram identificados 13 miRNAs diferencialmente expressos (regulados positivamente). A intersecção desses resultados revelou miRNAs que podem ser utilizados como biomarcadores sistêmicos para osteoartrite tibiotársica. A detecção do aumento na expressão destes miRNAs (eca-miR-15a, eca-miR-15b, eca-miR-17, eca-miR-18a, eca-miR-19b, eca-miR-20a, eca-miR-106a, eca-miR-106b, eca-miR-191, eca-miR-361-5p, eca-miR-450b-5p, eca-miR-451 e eca-miR-486-5p) no sangue venoso de equinos Quarto de Milha, indica que estes podem ser utilizados como biomarcadores para detecção precoce de osteoartrite tibiotársica em equinos de maneira menos invasiva e sistêmica.
Osteoarthritis (OA) is one of the major causes of morbidity in horses. It is characterized by articulation degeneration and bone proliferation, structures of the synovial membrane and joint capsule. The study of miRNA is an emerging field in determining the mechanisms that regulate inflammatory diseases, and can be present in models that might predict its effect on OA signaling pathways. Six athletic Quarter Horses were used, divided into two groups: Control Group (CT) - without any disease, and Osteoarthritis Group (OA) - diagnosed with subclinical traumatic primary tibiotarsal osteoarthritis. Venous blood and synovial fluid samples were collected to perform total RNA extraction using the “Alternative Protocol for Extraction of RNA from Cells Captured on LeukoLOCK Filters Using TRI Reagent®” kit (Thermo Fisher Scientific®). The microarray was performed with the kit “Affymetrix® miRNA 4.1 Array Strip” kit and “GeneAtlas Affymetrix®” equipment. Statistical analyzes were performed using the “Transcriptome Analysis Console (TAC)” program. When comparing groups, 42 miRNAs were identified differentially expressed in the venous blood of horses (36 upregulated and 6 down-regulated). In the synovial fluid, 13 differentially expressed (upregulated) miRNAs were identified. The intersection of these results revealed miRNAs that can be used as systemic biomarkers for tibiotarsal OA. The simultaneous detection of increased expression of these miRNAs (eca-miR-15a, eca-miR-15b, ecamiR-17, eca-miR-18a, eca-miR-19b, eca-miR-20a, eca-miR-106a, eca-miR-106b, eca-miR-191, eca-miR-361-5p, eca-miR-450b-5p, eca-miR-451 e eca-miR-486-5p) in venous blood of Quarter Horses, indicates that these molecules can be used as biomarkers for early detection of subclinical traumatic primary tibiotarsal osteoarthritis in horses in a less invasive and systemic way.
Osteoarthritis (OA) is one of the major causes of morbidity in horses. It is characterized by articulation degeneration and bone proliferation, structures of the synovial membrane and joint capsule. The study of miRNA is an emerging field in determining the mechanisms that regulate inflammatory diseases, and can be present in models that might predict its effect on OA signaling pathways. Six athletic Quarter Horses were used, divided into two groups: Control Group (CT) - without any disease, and Osteoarthritis Group (OA) - diagnosed with subclinical traumatic primary tibiotarsal osteoarthritis. Venous blood and synovial fluid samples were collected to perform total RNA extraction using the “Alternative Protocol for Extraction of RNA from Cells Captured on LeukoLOCK Filters Using TRI Reagent®” kit (Thermo Fisher Scientific®). The microarray was performed with the kit “Affymetrix® miRNA 4.1 Array Strip” kit and “GeneAtlas Affymetrix®” equipment. Statistical analyzes were performed using the “Transcriptome Analysis Console (TAC)” program. When comparing groups, 42 miRNAs were identified differentially expressed in the venous blood of horses (36 upregulated and 6 down-regulated). In the synovial fluid, 13 differentially expressed (upregulated) miRNAs were identified. The intersection of these results revealed miRNAs that can be used as systemic biomarkers for tibiotarsal OA. The simultaneous detection of increased expression of these miRNAs (eca-miR-15a, eca-miR-15b, ecamiR-17, eca-miR-18a, eca-miR-19b, eca-miR-20a, eca-miR-106a, eca-miR-106b, eca-miR-191, eca-miR-361-5p, eca-miR-450b-5p, eca-miR-451 e eca-miR-486-5p) in venous blood of Quarter Horses, indicates that these molecules can be used as biomarkers for early detection of subclinical traumatic primary tibiotarsal osteoarthritis in horses in a less invasive and systemic way.
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Palavras-chave
Articulação, Artropatias, Osteoartrite, Cavalo, microRNA, Articulation, Arthropathies, Osteoarthritis, Horse