Exercise training prevents the perivascular adipose tissue-induced aortic dysfunction with metabolic syndrome

Figures

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  Fig. 1. Effect of Exercise on tPVAT Mediated Aortic Function. Aortic total superoxide dismutase (SOD) activity (A, n=3–5), oxidative load measured by dihydroethidium (B) and coumarin boronate (C) with and without tPVAT incubation (n=5–8). Aortic nitric oxide (•NO) production in the presence or absence of tPVAT (D, n=8) was measured by 4-Amino-5-methylamino-2′,7′-difluorofluorescein diacetate (DAF-FM). Aortic relaxation to methacholine (MCh) in the presence or absence of tPVAT conditioned media in LZR (E, n=8) and OZR (F, n=8). Data are represented as Mean ± SD. For panels A-D, *p < 0.05, **p < 0.01, ***p < 0.001 compared to LZR or between groups connected by a bar. For panels E & F, *p < 0.05, **p < 0.01, ***p < 0.001 compared to LZR or OZR control without tPVAT, #p < 0.05 vs. LZR or OZR Ex without tPVAT.

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  Fig. 2. tPVAT Environment. qRT-PCR was used to assess tPVAT relative gene expression of phenotype indicators and immune cell markers (A, n=3), while immune-attracting cytokines (keratinocyte chemoattractant (KC)/human growth-regulated. oncogene (GRO), monocyte chemoattractant protein-1 (MCP-1), and lipocalin 1 were measured in tPVAT conditioned media (B, n=5). tPVAT production of •NO measured by fluorescent DAF-FM (C, n=8), oxidative load measured by dihydroethidium (D, n=5) and coumarin boronate (E, n=5), and relative SOD activity (F, n=4–6). Finally, gene expression of two subunits of the NOX2 enzyme (Gp91phox, p47phox) (G, n=3). Data are represented as Mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 compared to LZR or between groups connected by a bar. UCP-1, uncoupling protein-1; CD, cluster of differentiation; eNOS, endothelial nitric oxide synthase; Gch-1, GTP cyclohydrolase; SOD, superoxide dismutase; NOX2, NADPH oxidase 2 catalytic subunit (GP91); p47phox, NADPH oxidase 2 intracellular regulatory subunit.

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  Fig. 3. Exercise Enhancement of Proteasome Function. tPVAT samples were measure for proteasome protease activity at all three 20S active sites (A, n=6–8); LLVY (chymotrypsin-like), RLR (trypsin-like) and nLPnLD (caspase-like site). tPVAT samples were analyzed for levels of ubiquitin and the 20S proteasome beta-subunit (B, n=5). Ubiquitin-dependent 26S proteasome substrate degradation was measured in tPVAT samples by a linier ubiquitin chain-GFP substrate (C, n=6–8). Aortic relaxation was assessed in LZR aortic rings incubated with LZR tPVAT pre-treated with the proteasome inhibitor MG132 (D, n=4). Effects of proteasome inhibition in OZR Ex tPVAT on endothelial dependent dilation of healthy (donor) aortas (E, n=4). Data are represented as Mean ± SD. For panels A-C, *p < 0.05, **p < 0.01, ***p < 0.001 compared to LZR or between groups connected by a bar. For panels D and E, **p < 0.01 compared to LZR control tPVAT or OZR Ex tPVAT with MG132.

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  Fig. 4. tPVAT Cytokine Profile. tPVAT conditioned media was assessed by multi-plex array for anti-inflammatory (A, n=5), and pro- inflammatory cytokine levels (B, n=5). Data are represented as Mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 compared to LZR or between groups connected by a bar. IL, interleukin; HMW adiponectin, high molecular weight adiponectin; TNFα, tumor necrosis factor alpha; IFN-γ, interferon gamma; TSP-1, thrombospondin 1.

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  Fig. 5. Exercise Does Not Protect the Aorta Against tPVAT Derived TNFα Mediated Impairment. EDD of OZR-Ex aortas after incubation with either OZR Ex-tPVAT (n=5) or with a crossover treatment with OZR control tPVAT (n=5), or LZR control tPVAT (n=5). Data are represented as Mean ± SD. *p < 0.05, ***p < 0.001 compared to OZR-Ex aorta.

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  Fig. 6. Summary of main findings from the study.