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Murine hepatic proteome adaptation to high-fat diets with different contents of saturated fatty acids and linoleic acid : α-linolenic acid polyunsaturated fatty acid ratios

Kamila P. Liput

Kamila P. Liput

  • Department of Institute of Genetics and Animal Biotechnology of the Polish Academy of Sciences, JastrzębiecMagdalenka, Poland
  • Institute of Biochemistry and Biophysics of the Polish Academy of SciencesWarsaw, Poland
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Liput, Kamila P.
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Adam Lepczyński

Adam Lepczyński

Department of Physiology, Cytobiology and Proteomics, West Pomeranian University of TechnologySzczecin, Poland
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Lepczyński, Adam
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Ewa Poławska

Ewa Poławska

Department of Institute of Genetics and Animal Biotechnology of the Polish Academy of Sciences, JastrzębiecMagdalenka, Poland
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Poławska, Ewa
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Magdalena Ogłuszka

Magdalena Ogłuszka

Department of Institute of Genetics and Animal Biotechnology of the Polish Academy of Sciences, JastrzębiecMagdalenka, Poland
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Ogłuszka, Magdalena
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Rafał Starzyński

Rafał Starzyński

Department of Institute of Genetics and Animal Biotechnology of the Polish Academy of Sciences, JastrzębiecMagdalenka, Poland
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Starzyński, Rafał
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Paweł Urbański

Paweł Urbański

Department of Institute of Genetics and Animal Biotechnology of the Polish Academy of Sciences, JastrzębiecMagdalenka, Poland
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Urbański, Paweł
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Agata Nawrocka

Agata Nawrocka

Department of Institute of Genetics and Animal Biotechnology of the Polish Academy of Sciences, JastrzębiecMagdalenka, Poland
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Nawrocka, Agata
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Aneta Jończy

Aneta Jończy

Department of Institute of Genetics and Animal Biotechnology of the Polish Academy of Sciences, JastrzębiecMagdalenka, Poland
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Jończy, Aneta
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Dorota Pierzchała

Dorota Pierzchała

Maria Skłodowska-Curie National Research Institute of OncologyWarsaw, Poland
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Pierzchała, Dorota
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Chandra S. Pareek

Chandra S. Pareek

  • Department of Infectious and Invasive Diseases and Veterinary Administration, Institute of Veterinary Medicine, Faculty of Biological and Veterinary Sciences, Nicolaus Copernicus UniversityToruń, Poland
  • Division of Functional Genomics in Biological and Biomedical Research, Centre for Modern Interdisciplinary Technologies, Nicolaus Copernicus UniversityToruń, Poland
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Pareek, Chandra S.
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Marcin Gołyński

Marcin Gołyński

  • Department of Infectious and Invasive Diseases and Veterinary Administration, Institute of Veterinary Medicine, Faculty of Biological and Veterinary Sciences, Nicolaus Copernicus UniversityToruń, Poland
  • Division of Functional Genomics in Biological and Biomedical Research, Centre for Modern Interdisciplinary Technologies, Nicolaus Copernicus UniversityToruń, Poland
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Gołyński, Marcin
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Grzegorz Woźniakowski

Grzegorz Woźniakowski

  • Department of Infectious and Invasive Diseases and Veterinary Administration, Institute of Veterinary Medicine, Faculty of Biological and Veterinary Sciences, Nicolaus Copernicus UniversityToruń, Poland
  • Division of Functional Genomics in Biological and Biomedical Research, Centre for Modern Interdisciplinary Technologies, Nicolaus Copernicus UniversityToruń, Poland
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Woźniakowski, Grzegorz
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Urszula Czarnik

Urszula Czarnik

Department of Pig Breeding, Faculty of Animal Bio-Engineering, University of Warmia and Mazury in OlsztynOlsztyn, Poland
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Czarnik, Urszula
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Mariusz Pierzchała

Mariusz Pierzchała

Department of Institute of Genetics and Animal Biotechnology of the Polish Academy of Sciences, JastrzębiecMagdalenka, Poland
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Pierzchała, Mariusz
  
Aug 08, 2024
Murine hepatic proteome adaptation to high-fat diets with different contents of saturated fatty acids and linoleic acid : α-linolenic acid polyunsaturated fatty acid ratios's Cover Image
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Published Online: Aug 08, 2024
Page range: 427 - 441
Received: Jan 30, 2024
Accepted: Jul 29, 2024
DOI: https://doi.org/10.2478/jvetres-2024-0041
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© 2024 Kamila P. Liput et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1.

Experimental design to elucidate liver proteomic profile in mice fed four fat-varied diets. PUFA – polyunsaturated fatty acids; STD – standard diet; SFA – diet high in saturated fatty acids; 14 :1 – diet high in PUFA with this linoleic acid to α-linolenic acid ratio; 5 : 1 – diet high in PUFA with this linoleic acid to α-linolenic acid ratio; qPCR – real-time quantitative polymerase chain reaction; 2DE – two-dimensional electrophoresis. PDQuest Advanced 8.0.1 (Bio-Rad Laboratories, Hercules, CA, USA) software used for analysis of protein profiles. MALDI-TOF/TOF – matrix-assisted laser desorption/ionisation–time-of-flight tandem mass spectrometry
Experimental design to elucidate liver proteomic profile in mice fed four fat-varied diets. PUFA – polyunsaturated fatty acids; STD – standard diet; SFA – diet high in saturated fatty acids; 14 :1 – diet high in PUFA with this linoleic acid to α-linolenic acid ratio; 5 : 1 – diet high in PUFA with this linoleic acid to α-linolenic acid ratio; qPCR – real-time quantitative polymerase chain reaction; 2DE – two-dimensional electrophoresis. PDQuest Advanced 8.0.1 (Bio-Rad Laboratories, Hercules, CA, USA) software used for analysis of protein profiles. MALDI-TOF/TOF – matrix-assisted laser desorption/ionisation–time-of-flight tandem mass spectrometry

Fig. 2.

Effect on mice body weight of six months’ provision of experimental diets varied in fat content. Data are expressed as mean ± standard error of the mean, n = 10 mice per group. SEM – standard error of the mean; STD – standard diet; SFA – diet high in saturated fatty acids; 14 : 1 – diet high in polyunsaturated fatty acids (PUFA) with this linoleic acid to α-linolenic acid ratio; 5 : 1 – diet high in PUFA with this linoleic acid to α-linolenic acid ratio; * – significance difference at P-value < 0.05; ** – significant difference at P-value < 0.01; *** – significant difference at P-value < 0.001
Effect on mice body weight of six months’ provision of experimental diets varied in fat content. Data are expressed as mean ± standard error of the mean, n = 10 mice per group. SEM – standard error of the mean; STD – standard diet; SFA – diet high in saturated fatty acids; 14 : 1 – diet high in polyunsaturated fatty acids (PUFA) with this linoleic acid to α-linolenic acid ratio; 5 : 1 – diet high in PUFA with this linoleic acid to α-linolenic acid ratio; * – significance difference at P-value < 0.05; ** – significant difference at P-value < 0.01; *** – significant difference at P-value < 0.001

Fig. 3.

Final body weights of mice measured after six months’ provision of experimental diets varied in fat content and overnight fasting (A). Liver weights at the time of dissection (B). Ratios of liver weight to body weight (C). Data are expressed as mean ± standard deviation, n = 8 mice per group. STD – standard diet; SFA – diet high in saturated fatty acids; 14 : 1 – diet high in polyunsaturated fatty acids (PUFA) with this linoleic acid to α-linolenic acid ratio; 5 : 1 – diet high in PUFA with this linoleic acid to α-linolenic acid ratio; * – significant difference at P-value < 0.05; ** – significant difference at P-value < 0.01; *** – significant difference at P-value < 0.001
Final body weights of mice measured after six months’ provision of experimental diets varied in fat content and overnight fasting (A). Liver weights at the time of dissection (B). Ratios of liver weight to body weight (C). Data are expressed as mean ± standard deviation, n = 8 mice per group. STD – standard diet; SFA – diet high in saturated fatty acids; 14 : 1 – diet high in polyunsaturated fatty acids (PUFA) with this linoleic acid to α-linolenic acid ratio; 5 : 1 – diet high in PUFA with this linoleic acid to α-linolenic acid ratio; * – significant difference at P-value < 0.05; ** – significant difference at P-value < 0.01; *** – significant difference at P-value < 0.001

Fig. 4.

Blood parameters in mice after six months’ provision of experimental diets varied in fat content. Blood glucose level measured from tail blood (A). Mean red blood cell count measured from heart blood (B). Mean platelet volume measured in heart blood (C). STD – standard diet; SFA – diet high in saturated fatty acids; 14 : 1 – diet high in polyunsaturated fatty acids (PUFA) with this linoleic acid to α-linolenic acid ratio; 5 : 1 – diet high in PUFA with this linoleic acid to α-linolenic acid ratio; The plot shows the mean value, and the individual data points are presented in colours and shapes specific to each group; * – P-value < 0.05; ** – P-value < 0.01; *** – P-value < 0.001
Blood parameters in mice after six months’ provision of experimental diets varied in fat content. Blood glucose level measured from tail blood (A). Mean red blood cell count measured from heart blood (B). Mean platelet volume measured in heart blood (C). STD – standard diet; SFA – diet high in saturated fatty acids; 14 : 1 – diet high in polyunsaturated fatty acids (PUFA) with this linoleic acid to α-linolenic acid ratio; 5 : 1 – diet high in PUFA with this linoleic acid to α-linolenic acid ratio; The plot shows the mean value, and the individual data points are presented in colours and shapes specific to each group; * – P-value < 0.05; ** – P-value < 0.01; *** – P-value < 0.001

Fig. 5.

Differential murine liver protein spots after six months’ provision of experimental diets varied in fat contents. Spot numbers (SSP) and areas of protein spots with different expression levels between experimental groups correspond to the SSP number in Supplementary Table S3
Differential murine liver protein spots after six months’ provision of experimental diets varied in fat contents. Spot numbers (SSP) and areas of protein spots with different expression levels between experimental groups correspond to the SSP number in Supplementary Table S3

Fig. 6.

Differentially expressed murine liver proteins after six months’ provision of experimental diets varied in fat contents. The heatmap shows the increase (red) and the decrease (green) based on the comparison of the average spot intensity of the standard diet group (STD), group on a diet high in polyunsaturated fatty acids (PUFA) with 14 : 1 linoleic acid to α-linolenic acid ratio (14 : 1), group on a diet high in polyunsaturated fatty acids (PUFA) with 5 : 1 linoleic acid to α-linolenic acid ratio (5 : 1) and the saturated fatty acid diet (SFA) group. Lipid metabolism proteins (A); amino acid metabolism proteins (B); oxidative stress proteins (C); carbohydrate metabolism proteins (D); proteins regulating cell death pathway (E). HDL – high-density lipoprotein; ALB – albumin; HMGCS2 – 3-hydroxy-3-methylglutaryl-CoA synthase 2; ACAA2 – acetyl-CoA acyltransferase 2; BHMT – betaine-homocysteine S-methyltransferase; OAT – ornithine aminotransferase, mitochondrial; MAT1A – S-adenosylmethionine synthase isoform type-1; FTCD – formimidoyltransferase-cyclodeaminase; ALDH6A1 – aldehyde dehydrogenase family 6 member A1; HGD – homogentisate 1,2-dioxygenase; SUOX – sulfite oxidase, mitochondrial; INMT – indolethylamine N-methyltransferase; CA3 – carbonic anhydrase 3; EIF2S1 – eukaryotic translation initiation factor 2 subunit 1; PRDX6 – peroxiredoxin-6; ALDH1A1 – aldehyde dehydrogenase 1A1; COX6A1 – cytochrome c oxidase subunit 6A; FBP1 – fructose-1,6-bisphosphatase 1; PGAM1 – phosphoglycerate mutase 1; KHK – ketohexokinase; ENO1 – alpha-enolase; GALK1 – galactokinase; TKFC – triokinase/FMN cyclase; RGN – regucalcin; ANXA5 – annexin A5; FTL1 – ferritin light chain 1; HBB-B1 – haemoglobin subunit beta-1; * – significant difference at P-value < 0.05; ** – significant difference at P-value < 0.01; *** – significant difference at P-value < 0.001
Differentially expressed murine liver proteins after six months’ provision of experimental diets varied in fat contents. The heatmap shows the increase (red) and the decrease (green) based on the comparison of the average spot intensity of the standard diet group (STD), group on a diet high in polyunsaturated fatty acids (PUFA) with 14 : 1 linoleic acid to α-linolenic acid ratio (14 : 1), group on a diet high in polyunsaturated fatty acids (PUFA) with 5 : 1 linoleic acid to α-linolenic acid ratio (5 : 1) and the saturated fatty acid diet (SFA) group. Lipid metabolism proteins (A); amino acid metabolism proteins (B); oxidative stress proteins (C); carbohydrate metabolism proteins (D); proteins regulating cell death pathway (E). HDL – high-density lipoprotein; ALB – albumin; HMGCS2 – 3-hydroxy-3-methylglutaryl-CoA synthase 2; ACAA2 – acetyl-CoA acyltransferase 2; BHMT – betaine-homocysteine S-methyltransferase; OAT – ornithine aminotransferase, mitochondrial; MAT1A – S-adenosylmethionine synthase isoform type-1; FTCD – formimidoyltransferase-cyclodeaminase; ALDH6A1 – aldehyde dehydrogenase family 6 member A1; HGD – homogentisate 1,2-dioxygenase; SUOX – sulfite oxidase, mitochondrial; INMT – indolethylamine N-methyltransferase; CA3 – carbonic anhydrase 3; EIF2S1 – eukaryotic translation initiation factor 2 subunit 1; PRDX6 – peroxiredoxin-6; ALDH1A1 – aldehyde dehydrogenase 1A1; COX6A1 – cytochrome c oxidase subunit 6A; FBP1 – fructose-1,6-bisphosphatase 1; PGAM1 – phosphoglycerate mutase 1; KHK – ketohexokinase; ENO1 – alpha-enolase; GALK1 – galactokinase; TKFC – triokinase/FMN cyclase; RGN – regucalcin; ANXA5 – annexin A5; FTL1 – ferritin light chain 1; HBB-B1 – haemoglobin subunit beta-1; * – significant difference at P-value < 0.05; ** – significant difference at P-value < 0.01; *** – significant difference at P-value < 0.001

Western blot antibodies for identification of liver proteins in mice fed four fat-varied diets

Target protein UniProtKB accession number Gene symbol Molecular weight Dilution Host species Conjugate Supplier Catalogue number
Primary antibody
Ornithine aminotransferase P29758 Oat 49 kDa 1 : 500 mouse monoclonal Santa Cruz Biotechnology Dallas, TX, USA sc-376050
Glyceraldehyde-3-phosphate dehydrogenase P00355 GAPDH 36 kDa 1 : 10,000 rabbit polyclonal Abcam, Cambridge UK ab190304
Peroxiredoxin 6 P30041 PRDX6 25 kDa 1 : 400 mouse monoclonal Santa Cruz Biotechnology, Dallas, TX, USA sc-101522
Ferritin light chain P02792 FTL 21 kDa 1 : 1000 rabbit polyclonal Abcam, Cambridge UK ab69090
Secondary antibody
m-IgGκ BP-HRP mouse IgGκ light chain–binding protein conjugated to horseradish peroxidase (HRP) polyclonal horseradish peroxidase (HRP) conjugates Santa Cruz Biotechnology, Dallas, TX, USA sc-516102
Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) goat Jackson ImmunoResearch, Ely, UK 111-035-003
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