Cardiovascular Risk Factors and Differential Transcriptomic Profile of the Subcutaneous and Visceral Adipose Tissue and Their Resident Stem Cells

Figure 1. (A) Heat map of the gene expression in the subcutaneous fat of volunteers with normal weight versus patients with obesity; (B) Ratio of gene expression in subcutaneous fat of volunteers with normal weight versus patients with obesity. * p < 0.05; ** p > 0.01; and *** p < 0.001.

Figure 2. Line regression between expression of validated genes and body mass index (BMI). Correlations were determined by linear correlations. (A) Line regression between AGER and body mass index; (B) Line regression between ANGPT2 and body mass index; (C) Line regression between CD68 and body mass index; (D) Line regression between IFI30 and body mass index; (E) Line regression between NOCH3 and body mass index; (F) Line regression between SPP1 and body mass index; (G) Line regression between DLL4 and body mass index; (H) Line regression between PLIN2 and body mass index; (I) Line regression between VEGFA and body mass index; (J) Line regression between PINPLA2 and body mass index.

Figure 3. (A) Heat map of the gene expression in subcutaneous versus visceral fat; (B) Fold change of gene expression in subcutaneous versus visceral fat. * p < 0.05; ** p > 0.01; and *** p < 0.001.

Figure 4. Box-plot diagram showing adipose-derived stem cells (ASCs)-transcriptomic profile. (A) Differential mRNA expression of TF in SAT-ASC and VAT-ASC from diabetic with obesity and non-diabetic patients; (B) Differential mRNA expression of VEGFA in SAT-ASC and VAT-ASC from obese diabetic and non-diabetic patients; (C) Differential miR126 expression in SAT-ASC and VAT-ASC from obese diabetic and non-diabetic patients; (D) Differential miR145 expression in SAT-ASC and VAT-ASC from diabetic with obesity and non-diabetic patients. (* p < 0.05; ** p > 0.01; and *** p < 0.001).

Figure 5. TF and VEGF expression in in Subcutaneous or Visceral Derived ASCs from Diabetic/No Diabetic Patients with Obesity. (A) TF and VEGFA expression in ASCs obtained from SAT (S) or VAT (V) in obese patients (OB) in presence (DM) or absence (nDM) of diabetes. Western blot analysis of TF and VEGFA proteins in ASCs. To test for equal loading Western blots were reproved by β-actin; (B) Quantitative analysis of TF and VEGFA to β-actin relative levels.

Figure 6. (A) ASC microvesicles (MVs) secretion. Influence of obesity and diabetes on ASC derived MVs. Number of MVs measured by flow cytometry (Total, annexin-V positive, TF positive and annexin-V/TF positive MVs); (B) miR126 and miR145 relative gene expression in S-MVs and V-MVs. (* p < 0.05).

Figure 7. Effect of ASC-MVs in endothelial cell migration. Human microvascular endothelial (HMEC-1) cells treated with ASC conditioned medium, MVs depleted or MVs rich medium. (A) Images show the wound repair at 10 h; (B) Cell coverage of the wounded area at 10 h. Results are expressed as % of covered wounded area ± SEM. * p < 0.05.

Figure 8. Effect of ASCs-MVs on protein expression in protein endothelial cells. (A) TF and VEGFR2 protein expression in migrating endothelial cells treated with media with (+) or without (-) MVs obtained from 48 h cultured of subcutaneous (S) or visceral (V) ASCs from patients with obesity without diabetes (OB-nDM) or with diabetes (OB-DM); (B) Quantitative analysis of TF to β-actin relative levels; (C) Quantitative analysis of VEGFR2 to β-actin relative levels. (* p < 0.05).

Figure 9. Inflammatory and angiogenic pathways differentially affected in SAT versus VAT, in green downregulated and red up regulated.