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

Liput, Kamila P.; Lepczyński, Adam; Poławska, Ewa; Ogłuszka, Magdalena; Starzyński, Rafał; Urbański, Paweł; Nawrocka, Agata; Jończy, Aneta; Pierzchała, Dorota; Pareek, Chandra S.; Gołyński, Marcin; Woźniakowski, Grzegorz; Czarnik, Urszula; Pierzchała, Mariusz

doi:10.2478/jvetres-2024-0041

Figures & Tables

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

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

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

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

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

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

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

Figures & Tables

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

Fig. 5.

Fig. 6.

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

Paradigm

My account