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Hepatic microRNA modulation might be an early event to non-alcoholic fatty liver disease development driven by high-fat diet in male mice

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Abstract

Introduction

Metabolic alterations caused by an imbalance of macronutrient consumption are often related to the modulation of microRNAs (miRNAs), which could alter mRNAs expression profile and accelerate the development of non-alcoholic fatty liver disease (NAFLD).

Aims

This study aimed to investigate the contribution of miRNAs in modulating early stages of NAFLD in mice submitted to a high-fat diet (HFD).

Methods and results

Male Swiss mice, fed either a control diet or an HFD for 1, 3, 7, 15, 30, 56 days, were assessed for metabolic alterations, gene expression and NAFLD markers. A hepatocyte cell line was used to investigate the effects of miR-370 modulation on enzymes involved in β-oxidation. Body weight and adiposity were higher after 7 days of HFD. Fasting glucose and insulin increased after 3 and 7 days of HFD, respectively. While hepatic lipid content increased from the first day on, hepatic glycogen had a decrease after 3 days of HFD consumption. miR-370 and Let-7 expression increased with acute and chronic exposure to HFD, accompanied by carnitine palmitoyltransferase 1A (Cpt1a), acyl-CoA dehydrogenase very long chain (Acadvl) and protein kinase AMP-activated Catalytic Subunit 2 (Prkaa2) downregulation, while decreased miR-122 expression was accompanied by 1-acylglycerol-3-phosphate-O-acyltransferase (Agpat) upregulation after 56 days of HFD consumption, some of them confirmed by in vitro experiments. Despite fluctuations in TNFa and IL6 mRNA levels, molecular modulation was consistent with hepatic TG and NAFLD development.

Conclusion

Hepatic miR-370-122-Let7 miRNA modulation could be the first insult to NAFLD development, preceding changes in glycemic homeostasis and adiposity.

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Abbreviations

NAFLD:

Non-alcoholic fatty liver disease

MetSyn:

Metabolic syndrome

IR:

Insulin resistance

HFD:

High-fat diet

miRNA:

MicroRNA

DIO:

Diet-induced obesity

Gpam:

Glycerol-3-phosphate acyltransferase 1

Agpat:

1-Acylglycerol-3-phosphate-O-acyltransferase

Mogat:

2-Acylglycerol O-acyltransferase

Dgat:

Diglyceride acyltransferase

Cpt1a :

Carnitine palmitoyltransferase 1a

TG:

Triglycerides

CHOL:

Cholesterol

VLDL:

Very-low-density lipoprotein

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Funding

This study was supported by Grants from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, Brasil (CAPES, Finance Code 001), National Council for Scientific and Technological Development (CNPq # 310129/2016-8), and São Paulo Research Foundation (FAPESP # 2016/18321-6). All authors declared no competitive interests and thanks to Espaço da Escrita: Pró-Reitoria de Pesquisa: UNICAMP for the language services provided.

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Authors and Affiliations

  1. Laboratory of Metabolic Disorders, School of Applied Sciences, University of Campinas – UNICAMP, 1300, Pedro Zaccaria St, Limeira, São Paulo, 13484-350, Brazil

    Carolina Panzarin, Laís Angélica de Paula Simino, Mariana Camargo Silva Mancini, Leticia Martins Ignácio-Souza, Marciane Milanski, Márcio Alberto Torsoni & Adriana Souza Torsoni

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  1. Carolina Panzarin
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  2. Laís Angélica de Paula Simino
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  3. Mariana Camargo Silva Mancini
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Contributions

AST, LAPS, and CP carried out the study concept and design, data interpretation, and manuscript writing. CP, LASP, and MCM conducted the data acquisition and analyses. AST, MAT, LMIS, and MM did the critical review and provided funding for the study. All authors have approved the final version of the article.

Corresponding author

Correspondence to Adriana Souza Torsoni.

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The authors declare no competing interests.

Ethical approval

The authors state that all procedures comply with the ethical standards of the relevant national guides on the care and use of laboratory animals set forth by the Brazilian College for Animal Experimentation (COBEA) and has been approved by the Ethical Committee for Animal Use (CEUA), Protocol 4349-1, UNICAMP. This article does not contain any studies with human participants performed by any of the authors.

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The Laboratory of Metabolic Disorders belongs to the Obesity and Comorbidities Research Center (OCRC).

Supplementary Information

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11033_2021_7072_MOESM1_ESM.tiff

Supplementary file1 (TIFF 353 kb) Supplementary Fig. 1 Relative expression levels of hepatic inflammatory cytokines. Transcript levels of TNF (a) and IL-6 (b) from all groups. Values are mean ± SEM (n = 2–5). One-way ANOVA was used for multiple comparisons. Different letters indicate statistical significance between groups (p ≤ 0.05)

11033_2021_7072_MOESM2_ESM.tiff

Supplementary file2 (TIFF 142 kb) Supplementary Fig. 2 Western blotting of Cpt1 and Acadvl of hepatocytes transfected with Anti-miR-370

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Panzarin, C., Simino, L.A.d.P., Mancini, M.C.S. et al. Hepatic microRNA modulation might be an early event to non-alcoholic fatty liver disease development driven by high-fat diet in male mice. Mol Biol Rep 49, 2655–2666 (2022). https://doi.org/10.1007/s11033-021-07072-8

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