Aerobic exercise decreases the number and transcript expression of inflammatory M1 macrophages and CD8+ T cells in the epicardial adipose tissue of female pigs

Figures

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  Figure 1.. Analysis of differentially expressed genes and enrichment of KEGG pathways in the EAT from exercised-trained female pigs. (A) Volcano plot of up-regulated and down-regulated genes in exercise-trained versus sedentary pigs. (B) Identification of KEGG pathways significantly enriched (upregulated genes) or negatively enriched (downregulated genes) in exercise-trained pigs. The threshold set points for DEG were fold change > 2 and adjusted P value < 0.05. NS: not significant. Log2FC: log base 2-fold change. Log10: log base 10

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  Figure 2.. Distribution by treatment group and cell type of nuclei from occluded and non-occluded EAT of exercise-trained and sedentary female pigs. (A) UMAP plot of single nuclei from EAT of exercise-trained and sedentary female pigs showing the origins of nuclei in different treatment groups. (B) UMAP plot of single nuclei from EAT of female pigs showing seven major cell types from sedentary (occluded and nonoccluded) and exercised (occluded and non-occluded) tissue samples. (C) Bubble heatmap demonstrating expression levels of marker genes for each cell (nuclei) type. N: non-occluded; O: occluded; Ex: exercise trained; Sed: sedentary. APSC: adipocyte progenitor cells.

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  Figure 3.. Quantity and distribution of cell types across all treatment groups. (A) Barplot of the number of cell types in each treatment group. (B) Barplot of the percentage of cell types in each treatment group. (C) Batchwise distribution of each cell type. N: non-occluded; O: occluded; Ex: exercise trained; Sed: sedentary. APSC: adipocyte progenitor cells.

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  Figure 4.. Characterization of the cell number and gene signatures of macrophages and T cells in EAT in response to exercise and coronary artery occlusion. (A) Barplot the number of cells in each treatment group. (B) Barplot the percentage of cells in each treatment group. (C) Violin plot of the expression of selected marker genes in each cell type from EAT. (D) UMAP plot of T cell and macrophage clusters identified by unsupervised clustering. (E) UMAP plot of single nuclei from T cells and macrophages showing the origins of nuclei in different treatment groups. N: non-occluded; O: occluded; Ex: exercise trained; Sed: sedentary. STAB1: stabilin 1. CD163: CD163 molecule. SKAP1: Src kinase-associated phosphoprotein 1. NCALD: neurocalcin delta. TOX: thymocyte selection associated high mobility group box.

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  Figure 5.. Number and gene signature of CD8+ T cells in EAT by exercise and coronary artery occlusion treatment group. (A) Barplot of the CD8+ T cell number in EAT from each treatment group. (B) Violin plot of the expression of selected marker genes for identification of the CD8+ T cell population in EAT from each treatment group. (C) Box plot of AUCell scores of T cells from EAT of each treatment group expressing genes in the tumor necrosis factor (TNF) superfamily. N: non-occluded; O: occluded; Ex: exercise trained; Sed: sedentary. CD8A: CD8A molecule. CCL5: C-C motif chemokine ligand 5. CD2: CD2 molecule. GZMH: granzyme H. TGFBR2: transforming growth factor receptor 2. AUCell uses the Area Under the Curve to determine if a subset of the input gene set is enriched in the expressed genes for a given cell type. NS: not significant. * P < 0.05. *** P < 0.001.

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  Figure 6.. Functional analysis of macrophage populations in EAT in response to exercise and coronary artery occlusion status. (A) UMAP plot of macrophages classified by respective marker genes. (B) Barplot of the number and percentage of M0, M1, and M2 macrophages in each treatment group. (C) Violin plot showing the expression level of selected marker genes in each macrophage class. (D) Box plot of AUCell scores of macrophages from EAT of each treatment group expressing genes in the tumor necrosis factor (TNF) superfamily. N: non-occluded; O: occluded; Ex: exercise trained; Sed: sedentary. CD209: CD209 molecule. TGFB1: transforming growth factor beta 1. CD80: CD80 molecule. IRF5: interferon regulatory factor 5. PPARG: peroxisome proliferator-activated receptor gamma

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  Figure 7.. Cell-to-cell communications between immune and non-immune cells in epicardial adipose tissue. (A) Dot plot of the number of significant interactions between cell types. Circle color correlates to the probability of cell-cell interaction. Circle size correlates to the number of cell-cell interactions. (B) Heat map of the crosstalk between immune cells, adipocytes, and APSCs in EAT. Communication strength for each interaction, as well as the sender and receiver, are shown for each cell-to-cell interaction. N_Ex: occluded exercise; N_Sed: occluded sedentary O_Ex: occluded exercise; O_Sed: occluded sedentary. ADI: adipocytes. APSC: adipocyte stem cells. MAC: macrophages. TC: T cells. See the abbreviation list for gene names.