Mesenchymal Stem/Stromal Cells Reverse Adipose Tissue Inflammation in Pigs with Metabolic Syndrome and Renovascular Hypertension

Figures

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  Figure 1. Schematic of the experimental protocol and treatment timeline.

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  Figure 2. MSCs downregulate visceral adipose tissue accumulation. (A) Abdominal CT axial images with post-imaging processing highlighting subcutaneous (red) and visceral (yellow) fat. (B) Visceral fat fraction was augmented in MetS and MetS + RAS, but not in MetS + RAS + MSC. (C) The subcutaneous fat fraction was increased in all 3 experimental groups relative to Lean. * p < 0.05 vs. Lean. MetS, metabolic syndrome; RAS, renal artery stenosis; MSC, mesenchymal stem/stromal cells.

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  Figure 3. MSCs attenuate adipose tissue fibrosis and hypertrophy. Representative images of trichrome-stained (blue) perirenal (A) and subcutaneous (B) fat (10× magnification). Pink: eosin counterstain. (C) Perirenal fat fibrosis increased in MetS + RAS but was attenuated by the delivery of MSCs. (D) Subcutaneous fat fibrosis was only higher in MetS + RAS vs. MetS. (E) Adipocyte cross-sectional area was higher in the perirenal fat of MetS + RAS vs. Lean, suggesting hypertrophy, but decreased in MetS + RAS + MSC. (F) In the subcutaneous fat, MetS had smaller adipocytes compared to Lean and MetS + RAS. * p < 0.05 vs. Lean; † p < 0.05 vs. MetS; $ p < 0.05 vs. MetS + RAS. MetS, metabolic syndrome; RAS, renal artery stenosis; MSC, mesenchymal stem/stromal cells.

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  Figure 4. MSCs downregulate perirenal adipose tissue inflammation. (A) Representative immunofluorescence images (40×) showing the expression of MCP-1 (red), TNF-a (green), and IL-6 (red) in pig perirenal fat. MCP-1 (B), TNF-a (C), and IL-6 (D) were upregulated in MetS + RAS relative to Lean and MetS but were downregulated in MetS + RAS + MSC. * p < 0.05 vs. Lean; † p < 0.05 vs. MetS; $ p < 0.05 vs. MetS + RAS. MetS, metabolic syndrome; RAS, renal artery stenosis; MSC, mesenchymal stem/stromal cell.

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  Figure 5. MSCs downregulate subcutaneous adipose tissue inflammation. (A) Representative immunofluorescence staining images (40×) showing the expression of MCP-1 (red), TNF-a (green), and IL-6 (red) in pig subcutaneous fat. MCP-1 (B) and TNF-a (C) were upregulated in MetS + RAS vs. Lean. MCP-1 significantly decreased in MetS + RAS + MSC and TNF-a tended to decrease as well. (D) IL-6 expression was not elevated in MetS or MetS + RAS vs. Lean. Nevertheless, MSCs reduced IL-6 expression compared to MetS + RAS. * p < 0.05 vs. Lean; † p < 0.05 vs. MetS; $ p < 0.05 vs. MetS + RAS. MetS, metabolic syndrome; RAS, renal artery stenosis; MSC, mesenchymal stem/stromal cell.

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  Figure 6. MSCs downregulate proinflammatory cytokine gene expression. (A,C,E) MCP-1 and IL-6 gene expression was upregulated in the MetS + RAS perirenal fat relative to Lean, but MSC treatment attenuated this effect. A slight elevation of TNF-a gene expression (p = 0.09) in the perirenal fat of MetS + RAS did not achieve statistical significance. (B,D,F) In the subcutaneous fat, no differences in MCP-1 or TNF-a gene expression were observed among the four groups. However, IL-6 mRNA expression was reduced in the MetS + RAS pigs, while in the MetS + RAS + MSC group, IL-6 levels trended lower relative to the Lean group (p = 0.08) but did not reach statistical significance. * p < 0.05 vs. Lean; † p < 0.05 vs. MetS. MetS, metabolic syndrome; RAS, renal artery stenosis; MSC, mesenchymal stem cells.

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  Figure 7. MSCs alter perirenal and subcutaneous anti-inflammatory and adipogenic gene expression. (A) Perirenal fat showed upregulated TSG-6 expression in MetS and MetS + RAS + MSC relative to Lean. (B) In contrast, subcutaneous fat showed downregulated TSG-6 expression in MetS and MetS + RAS relative to Lean. MetS + RAS + MSC tended to be lower than Lean, but this did not reach statistical significance (p = 0.06). C/EBPa expression increased only in the perirenal fat of MetS + RAS + MSC pigs relative to Lean (C) and in the subcutaneous fat of MetS + RAS relative to Lean and MetS (D). (E) Perirenal fat C/EBPb expression was upregulated in MetS relative to Lean and in MetS + RAS + MSC relative to Lean and MetS + RAS. (F) Subcutaneous fat expression of C/EBPb was upregulated in MetS + RAS relative to Lean and MetS. PPARy was upregulated in the perirenal fat of MetS + RAS relative to Lean (G) but was unchanged in the subcutaneous fat of all pig groups (H). * p < 0.05 vs. Lean; † p < 0.05 vs. MetS; $ p < 0.05 vs. MetS + RAS. MetS, metabolic syndrome; RAS, renal artery stenosis; MSC, mesenchymal stem cells.