Tesis
Tratamento in vitro e in vivo com laser de baixa intensidade em camundongos mdx = In vitro and in vivo low level laser therapy treatment in mdx mice
In vitro and in vivo low level laser therapy treatment in mdx mice
Registro en:
MACEDO, Aline Barbosa. Tratamento in vitro e in vivo com laser de baixa intensidade em camundongos mdx = In vitro and in vivo low level laser therapy treatment in mdx mice. 2016. 1 recurso online ( 107 p.). Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia, Campinas, SP.
Autor
Macedo, Aline Barbosa, 1986-
Institución
Resumen
Orientador: Elaine Minatel Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia Resumo:A distrofia muscular de Duchenne (DMD) e uma doenca letal que afeta 1 em cada 3.500 criancas do sexo masculino nascidas vivas. As especies reativas de oxigenio (EROs) e a resposta inflamatoria exacerbada desempenham um papel fundamental na patogenese da DMD. Evidencias sugerem que a laserterapia de baixa intensidade reduz a producao de EROs, diminui o processo inflamatorio e aumenta a producao de antioxidantes. Com essa perspectiva em mente, o presente estudo investigou os efeitos in vitro e in vivo da LTBI nas fibras musculares distroficas de camundongos mdx, modelo experimental da DMD. As culturas primarias de celulas musculares mdx foram irradiadas uma unica vez com laser e analisadas apos 24 e 48 horas. Enquanto, no estudo in vivo, o musculo quadriceps distrofico foi tratado com laser por tres vezes por semana, durante duas semanas. Ambos os estudos consideraram o camundongo C57BL/10 como controle. O comprimento de onda utilizado foi de 830 nm e 5 J/cm2 de fluencia. A LTBI promoveu: (1) aumento na viabilidade e proliferacao celular distrofica, observadas atraves dos ensaios de Vermelho neutro, MTT e aumento no diametro das fibras musculares; (2) favoreceu a regeneracao muscular, reduzindo a area de fibras em degeneracao, os niveis de MyoD e Atrogin 1/MAFbx e aumentando os niveis de MyHC e VEGF; (3) aumento transitorios na concentracao de calcio intracelular; (4) reducao do estresse oxidativo, diminuindo a producao de H2O2, do marcador de peroxidacao lipidica (4-HNE), deteccao de anions superoxidos (DHE) e do sistema antioxidante (GSH, GR, GPx, SOD e Catalase); (5) reducao da resposta inflamatoria, area de inflamacao, area de infiltrado de macrofagos, niveis de TNF-?e NF-kB. Estes resultados sugerem que a LTBI pode constituir uma potencial terapia alternativa para ser associada com outros tratamentos farmacologicos no tratamento da DMD Abstract: Duchenne muscular dystrophy (DMD) is a lethal disease that affects 1 in every 3,500 male children born alive. Reactive oxygen species (ROS) and exacerbated inflammatory response play a key role in the pathogenesis of DMD. Evidence suggests that low level laser therapy (LLLT) reduces ROS production, decreases the inflammatory process and increase the production of antioxidants. With this perspective in mind, the present study investigated the in vitro and in vivo effects of LLLT in the dystrophic muscle fibers of mdx mice, an experimental model of DMD. Primary cultures of mdx muscle cells were irradiated once with laser and analyzed after 24 and 48 hours. While the in vivo study, the dystrophic quadriceps muscle was treated with laser for three times per week for two weeks. Both studies found the mouse C57BL/10 as a control. The wavelength used was 830 nm and 5 J/cm² fluence. LLLT promoted: (1) increased cell viability and proliferation in dystrophic observed by Neutral red, MTT assays and an increase in the diameter of muscle fibers; (2) favored the muscle regeneration, reducing the degeneration fibers area, the levels of MyoD and atrogin 1/MAFbx and increasing levels of MyHC and VEGF; (3) increase in temporary intracellular calcium concentration; (4) reduction of oxidative stress by decreasing the production of H2O2, the marker of lipid peroxidation (4-HNE), superoxide anions detection (DHE) and the antioxidant system (GSH, GR, GPx, SOD and Catalase); (5) decrease the inflammatory response, inflammatory area, macrophage infiltrate area, TNF-? and NF-kB levels. These results suggest that LLLT may constitute a potential alternative therapy to be associated with other pharmacological treatments for the treatment of DMD Doutorado Anatomia Doutora em Biologia Celular e Estrutural CNPQ CAPES
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