Proteoma miocárdico de ratos obesos por dieta Ocidental com disfunção cardíaca
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Data
2019-02-26
Autores
Vileigas, Danielle Fernandes
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
A obesidade é uma doença metabólica complexa considerada uma pandemia global e associada à alta incidência de doença cardiovascular. O excesso de tecido adiposo pode promover mal adaptação que resulta em alterações na estrutura e função do coração; no entanto, os mecanismos não estão totalmente elucidados. A proteômica pode fornecer uma compreensão mais profunda do processo fisiopatológico e contribuir para a identificação de novos potenciais alvos terapêuticos. Portanto, o objetivo deste estudo foi avaliar a expressão proteica miocárdica em ratos saudáveis e obesos por dieta Ocidental, empregando duas abordagens proteômicas, para melhor compreender a rede de mecanismos inerentes à disfunção cardíaca na obesidade. Ratos Wistar foram distribuídos em dois grupos: controle (C, n = 13; dieta controle) e obeso (Ob, n = 13; dieta Ocidental) alimentados por 41 semanas. A obesidade foi determinada pelo índice de adiposidade. A função cardíaca foi avaliada pelo ecocardiograma e análise do músculo papilar isolado. A proteômica foi baseada em eletroforese em gel bidimensional (2-DE) juntamente com espectrometria de massa (LC-MS/MS) e cromatografia-líquida em nanofluxo com espectrometria de massa em tandem (nanoLC-MS/MS) seguida de quantificação label-free. Ratos obesos apresentaram aumento do índice de adiposidade e disfunção cardíaca sistólica e diastólica comparados aos controles. Um total de 82 proteínas miocárdicas foram identificadas como diferencialmente expressas entre os grupos C e Ob usando duas abordagens proteômicas distintas, sendo que 43 e 39 apresentaram maiores e menores níveis de expressão, respectivamente, nos animais Ob. Estas proteínas estão envolvidas em importantes processos biológicos, incluindo a beta-oxidação dos ácidos graxos na mitocôndria e peroxissoma, homeostase lipídica intracelular, processo metabólico de glicose e aminoácidos, ciclo do ácido tricarboxílico, fosforilação oxidativa, defesas antioxidantes, função contrátil e regulação do trânsito de cálcio intracelular. Os dados proteômicos indicaram elevada expressão de várias proteínas do citoesqueleto, assim como diminuição nos níveis de proteínas do complexo proteassoma no miocárdio de ratos Ob, as quais não têm sido previamente associadas à disfunção cardíaca na obesidade. Além disso, a combinação de ambas as técnicas proteômicas pode ser relevante para aumentar o mapa proteômico cardíaco. Em conclusão, os achados deste estudo revelaram diversas alterações no proteoma miocárdico de ratos obesos, possibilitando novos insights sobre os mecanismos envolvidos no comprometimento funcional cardíaco devido à obesidade e possíveis novos alvos terapêuticos.
Obesity is a complex metabolic disease considered a global pandemic and associated with high incidence of cardiovascular disease. The excess of adipose tissue may promotes maladaptation that result in alterations in structure and function of the heart; however, the mechanisms are not fully elucidated. Proteomics may provide a deeper understanding into the pathophysiological process and contribute to the identification of new potential therapeutic targets. Thus, the aim of this was evaluate the myocardial protein expression in healthy and obese rats, employing two proteomic approaches to better comprehend the network of mechanisms inherent to cardiac dysfunction in obesity. Male Wistar rats were distributed into two groups: control (C, n=13; standard diet) and obese (Ob, n=13; Western diet) fed for 41 weeks. The obesity was determined by adipose index. Cardiac function was evaluated by echocardiogram and isolated papillary muscle analysis. The proteomics was based on two-dimensional gel electrophoresis (2-DE) along with mass spectrometry identification (LC-MS/MS) and nano-liquid chromatography with tandem mass spectrometry (nanoLC-MS/MS) followed by label-free quantification. Obese rats showed increased adiposity index and systolic and diastolic cardiac dysfunction. A total of 82 myocardial proteins was identified as differentially expressed between C and Ob groups using two proteomic strategies, being 43 up- and 39 down-regulated by obesity. These proteins are involved in important biological processes including mitochondrial and peroxisomal fatty acid beta-oxidation, intracellular lipid homeostasis, metabolic process of glucose and amino acid, tricarboxylic acid cycle, oxidative phosphorylation, antioxidant defenses, contractile function and regulation of calcium transient. Interestingly, the proteomic data indicated high and low expression of several cytoskeletal and proteasome complex proteins, respectively, in obese rat myocardium, which have not previously been associated with cardiac dysfunction in obesity. Furthermore, the combination of both 4 proteomic techniques may be relevant to increase the cardiac proteomic map. In conclusion, the proteomic findings revealed diverse alterations in myocardial proteome from obese rats, enabling novel insights concerning the mechanisms involved in cardiac functional impairment due to obesity and possible new therapeutic targets.
Obesity is a complex metabolic disease considered a global pandemic and associated with high incidence of cardiovascular disease. The excess of adipose tissue may promotes maladaptation that result in alterations in structure and function of the heart; however, the mechanisms are not fully elucidated. Proteomics may provide a deeper understanding into the pathophysiological process and contribute to the identification of new potential therapeutic targets. Thus, the aim of this was evaluate the myocardial protein expression in healthy and obese rats, employing two proteomic approaches to better comprehend the network of mechanisms inherent to cardiac dysfunction in obesity. Male Wistar rats were distributed into two groups: control (C, n=13; standard diet) and obese (Ob, n=13; Western diet) fed for 41 weeks. The obesity was determined by adipose index. Cardiac function was evaluated by echocardiogram and isolated papillary muscle analysis. The proteomics was based on two-dimensional gel electrophoresis (2-DE) along with mass spectrometry identification (LC-MS/MS) and nano-liquid chromatography with tandem mass spectrometry (nanoLC-MS/MS) followed by label-free quantification. Obese rats showed increased adiposity index and systolic and diastolic cardiac dysfunction. A total of 82 myocardial proteins was identified as differentially expressed between C and Ob groups using two proteomic strategies, being 43 up- and 39 down-regulated by obesity. These proteins are involved in important biological processes including mitochondrial and peroxisomal fatty acid beta-oxidation, intracellular lipid homeostasis, metabolic process of glucose and amino acid, tricarboxylic acid cycle, oxidative phosphorylation, antioxidant defenses, contractile function and regulation of calcium transient. Interestingly, the proteomic data indicated high and low expression of several cytoskeletal and proteasome complex proteins, respectively, in obese rat myocardium, which have not previously been associated with cardiac dysfunction in obesity. Furthermore, the combination of both 4 proteomic techniques may be relevant to increase the cardiac proteomic map. In conclusion, the proteomic findings revealed diverse alterations in myocardial proteome from obese rats, enabling novel insights concerning the mechanisms involved in cardiac functional impairment due to obesity and possible new therapeutic targets.
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Palavras-chave
Obesidade, Dieta rica em gordura e açúcar, Função cardíaca, Proteômica shotgun e quantificação label-free, Eletroforese em gel bidimensional, Proteoma miocárdico, Ratos, Obesity, High-fat high-sugar diet, Cardiac function, Shotgun and labelfree quantification, Two-dimensional electrophoresis, Myocardial proteome, Rats