Avaliação da relação entre ritmos biológicos e fisiologia dos lisossomos e sua participação em modelo de doença lisossômica

• Ano de defesa: 2016
• Autor(a) principal: Oliveira, Ana Carolina Barris de [UNIFESP]
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de São Paulo (UNIFESP)
• Palavras-chave em Português: Redox state; Lysosomes; Lisossomos; Ritmos biológicos; Sistema de temporização; Doença de fabry; Estado redox
• Link de acesso: https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=3615253; http://repositorio.unifesp.br/handle/11600/47692

The clock system is responsible for the orchestration observed in several physiological, cellular and molecular processes. This regulation is controlled by a molecular machinery that allows the ideal synchronization of each tissue to the organism and this control presents plasticity that ensures the adaptation of the system to environmental changes. Lysosomes, organelles responsible for cellular digestion, present circadian activities and the role of the clock system is being explored in Lysosomal Storage Diseases, in which oxidative stress has a participation in their pathophysiologies and is regulated by the clock system. Despite these information, the relation between lysosomes and clock system in poorly understood. Thus, the aim of this work was to evaluate consequences of lysosomal malfunction, induced by drug or genetic disorder, in clock system and redox state. Rat fibroblasts with the gene Bmal1 conjugated to luciferase were treated with lysosomal inhibitors at the peak and trough of Bmal1 expression for Bmal1 expression and superoxide dismutase (SOD1) analysis; rat cardiomyocytes were treated at the peak of expression of Bmal1 for clock genes expression and autophagy-related proteins analysis and; control individuals and Fabry Disease (FD) patients? fibroblasts were used for clock genes and antioxidant enzymes genes expression evaluation, SOD1 and LAMP-2 protein expression and verify if the GLA gene, mutated in FD, and ?-galactosidase A enzyme (?-gal A), deficient if FD, present circadian oscillation. Results obtained indicated that the lysosomal inhibition by drugs promotes phase advance or delay and period alterations in Bmal1 expression, depending on the moment of treatment ? peak or trough of Bmal1 expression; besides acute modifications in Bmal1 and Per1, clock genes, and, phosphorylated-AMPK and LC-3 expression, proteins involved in autophagy. FD patients? fibroblasts presented altered expression of clock genes, especially from the positive arm of regulation (BMAL1 and CLOCK); GLA gene presented circadian oscillation and ?-gal A seemed to be influenced by the clock system. Redox state regulation was altered by lysosomal inhibition by drugs, in FD patients? fibroblasts the antioxidant enzymes genes and SOD1 did not present compromised clock system regulation but the protein LAMP-2 had alterations of its rhythmicity in FD patients? cells. Based on all these analysis, we suggest that lysosomal inhibition, acute or chronic, promotes alterations in the molecular regulation of the clock system likely related to the increase of cytosolic calcium. Therefore, we concluded that lysosomes have a role in the regulation of clock system and redox state, and hypothesize that this may happen through calcium mobilization from the interior of these organelles to the cytosol.