Unraveling Hepatic Metabolomic Profiles and Morphological Outcomes in a Hybrid Model of NASH in Different Mouse Strains

Abstract

Nonalcoholic fatty liver disease (NAFLD) encompasses nonalcoholic steatohepatitis (NASH) and affects 25% of the global population. Although a plethora of experimental models for studying NASH have been proposed, still scarce findings regarding the hepatic metabolomic/molecular profile. In the present study, we sought to unravel the hepatic metabolomic profile of mice subjected to a hybrid model of NASH, by combining a Western diet and carbon tetrachloride administration, for 8 weeks, in male C57BL/6J and BALB/c mice. In both mouse strains, the main traits of NASH—metabolic (glucose intolerance profile), morphologic (extensive microvesicular steatosis and fibrosis, lobular inflammation, and adipose tissue-related inflammation/hypertrophy), and molecular (impaired Nrf2/NF-κB pathway dynamics and altered metabolomic profile)—were observed. The hepatic metabolomic profile revealed that the hybrid protocol impaired, in both strains, the abundance of branched chain-aromatic amino acids, carboxylic acids, and glycosyl compounds, that might be linked to the Nrf2 pathway activation. Moreover, we observed a strain-dependent hepatic metabolomic signature, in which the tricarboxylic acid metabolites and pyruvate metabolism were dissimilarly modulated in C57BL/6J and BALB/c mice. Thus, we provide evidence that the strain-dependent hepatic metabolomic profile might be linked to the distinct underlying mechanisms of NASH, also prospecting potential mechanistic insights into the corresponding disease.

Comprehensive Strategy for Identifying Extracellular Vesicle Surface Proteins as Biomarkers for Non-Alcoholic Fatty Liver Disease

“…CD68 is a well-established marker for identifying macrophages. Increased liver infiltration with CD68+ macrophages is related to liver fibrosis [77]. In addition, CD68 expression has been found on circulating EVs in hypertensive rats [78].…”

Comprehensive Strategy for Identifying Extracellular Vesicle Surface Proteins as Biomarkers for Non-alcoholic Fatty Liver Disease

“…CD68 is a well-established marker for identify macrophages. Increased liver infiltration with CD68+ macrophages are related to liver fibrosis [72]. In addition, CD68 expression has been found on circulating EVs in hypertensive rats [73].…”

Abordagem metabolômica do fígado de bovinos como busca de biomarcadores para mitigar a emissão de metano

“…Os espectros foram adquiridos para RMN de prótons usando a sequência de pulso PRESAT para a supressão do sinal de água residual, 32 K pontos de dados, com uma largura espectral de 14 ppm, com um tempo de aquisição de 4,0 s, foram adquiridos com um ganho fixo do receptor de 26 e um atraso de reciclagem de 20 s (5*T1), varreduras fictícias de 2 e um acúmulo de 128 transientes por 52 minutos e 12 segundos por análise (BACIL et al, 2023).…”

“…de compostos polares e apolares, utilizou-se a metodologia descrita porBacil et al (2023). As amostras de fígado bovino dos 24 animais armazenadas em -80 °C foram colocadas em tubo de homogeneização livres de RNase e DNase de 2ml contendo grânulos de cerâmica com diâmetro de 1,4 mm (matriz D, MP Biomedicals, EUA).Primeiramente 200 mg de amostras de fígado bovino foram utilizadas para a extração com 300 μL de clorofórmio, metanol, água destilada (1:2:1 v/v), a solução de extração foi adicionada a cada tubo e os tecidos foram então homogeneizados em um homogeneizador FastPrep-24 (MP Biomedicals, EUA) em dois ciclos a 4 m/s por 30 s com intervalo de pausa de 30 s entre os ciclos.Após a homogeneização das amostras adicionou-se 200 μL de clorofórmio para separação de fases e centrifugou-se a 4 ºC e 1500×g por 15 min, nesta etapa houve a separação das fases correspondendo às amostras polares e apolares, a fase aquosa hidroalcóolica foi coletada e concentrada em um SpeedVac Vacuum Concentrator (Thermo Fisher Scientific, EUA) e armazenada a -80 ºC até as análises metabolômicas.…”