Journal: Experimental physiology

Article Title: Aortic Dysfunction in Metabolic Syndrome Mediated by Perivascular Adipose Tissue TNFα and NOX2 Dependent Pathway

doi: 10.1113/EP086818

Figure Lengend Snippet: Relative gene expression of OZR tPVAT compared to LZR tPVAT for A) phenotype and immune markers, B) oxidative and inflammatory genes, and C) and anti-inflammatory and oxidative defense markers. Data expressed as Mean±SD, *denotes significant difference in OZR vs. LZR, minimum of 2-fold change and t-test p<0.05, n=3. UCP-1, uncoupling protein-1; CD, cluster of differentiation; NOX2, NADPH oxidase 2 catalytic subunit (GP91); p47phox, NADPH oxidase 2 intracellular regulatory subunit; TNF, tumor necrosis factor; CCL5, Chemokine (C-C motif) ligand 5; IL, interleukin; TSP-1, thrombospondin 1; IFN-γ, interferon gamma; CCR, C-C motif chemokine receptor; AdipoQ, adiponectin; NFR2, nuclear factor (erythroid 2)-like 2; Keap1, kelch-like ECH associated protein 1, SOD, superoxide dismutase; GSR, glutathione reductase; CAT, catalase.

Article Snippet: Gene Expression 50mg sections of tPVAT were incubated at 37°C in physiological HEPES buffer (43.7 mM NaCl, 80 mM KCl, 1.17 mM MgSO 4 •7H 2 O, 1.6 mM NaH 2 PO 4 , 18 mM NaHCO 3 , 0.03 mM EDTA, 5.5 mM glucose, 5 mM HEPES) or HEPES buffer containing 4μM TNFα neutralizing antibody (TNFα-AB, Catalog #: AF-510-NA, R&D systems) at a ratio of 200μg/mL.

Techniques: Gene Expression

Journal: Experimental physiology

Article Title: Aortic Dysfunction in Metabolic Syndrome Mediated by Perivascular Adipose Tissue TNFα and NOX2 Dependent Pathway

doi: 10.1113/EP086818

Figure Lengend Snippet: The secretion profile of tPVAT exudate was assessed for A & B) immuno-attractive cytokines, inflammatory cytokines, and C & D) anti-inflammatory cytokines (n=5). Data expressed as Mean±SD *denotes a statistically significant change in OZR vs. LZR determined by t-test, p<0.05. KC/GRO, chemokine (C-C motif) ligand 1; MCP-1, monocyte chemoattractant protein-1; TNFα, tumor necrosis factor alpha; IL, interleukin; IFN-γ, interferon gamma; TSP-1, thrombospondin 1; HMW adiponectin, high molecular weight adiponectin

Article Snippet: Gene Expression 50mg sections of tPVAT were incubated at 37°C in physiological HEPES buffer (43.7 mM NaCl, 80 mM KCl, 1.17 mM MgSO 4 •7H 2 O, 1.6 mM NaH 2 PO 4 , 18 mM NaHCO 3 , 0.03 mM EDTA, 5.5 mM glucose, 5 mM HEPES) or HEPES buffer containing 4μM TNFα neutralizing antibody (TNFα-AB, Catalog #: AF-510-NA, R&D systems) at a ratio of 200μg/mL.

Techniques: High Molecular Weight

Journal: Experimental physiology

Article Title: Aortic Dysfunction in Metabolic Syndrome Mediated by Perivascular Adipose Tissue TNFα and NOX2 Dependent Pathway

doi: 10.1113/EP086818

Figure Lengend Snippet: A) Effect of ex-vivo TNFα-AB treatment on activation of NF-κB in tPVAT shown as the ratio of phosphor:total (n=6), and B) the effect on tPVAT gene expression compared to untreated OZR tPVAT (n=3). Following the ex-vivo treatment of OZR tPVAT with TNFα-AB, NOX2ds-TAT or a combination C) ROS production was assessed in tPVAT and aorta incubated with tPVAT (n=8, repeated measures were used to assess the effect of treatment on tPVAT ROS and separately on the effect of aortic ROS, separation of analysis shown by dashed line). The effect of ex-vivo TNFα-AB treatment on OZR-tPVAT mediated D) NO production and E&F) EDD in OZR aorta and the crossover experiment in LZR aorta (n=5–8). Data expressed as Mean±SD. ^denotes significant effect of treatment compared to control assessed by repeated measures ANOVA, p<0.05. Pe, Phenylephrine; MCh, Methacholine; TNF, tumor necrosis factor; NOX2, NADPH oxidase 2 catalytic subunit (GP91); p47phox, NADPH oxidase 2 intracellular regulatory subunit; TSP-1, thrombospondin 1; IL, interleukin; IFN-γ, interferon gamma; NFR2, nuclear factor (erythroid 2)-like 2; AdipoQ, adiponectin MMP, matrix metalloproteinase; TIMP, tissue inhibitor of metalloproteinase.

Article Snippet: Gene Expression 50mg sections of tPVAT were incubated at 37°C in physiological HEPES buffer (43.7 mM NaCl, 80 mM KCl, 1.17 mM MgSO 4 •7H 2 O, 1.6 mM NaH 2 PO 4 , 18 mM NaHCO 3 , 0.03 mM EDTA, 5.5 mM glucose, 5 mM HEPES) or HEPES buffer containing 4μM TNFα neutralizing antibody (TNFα-AB, Catalog #: AF-510-NA, R&D systems) at a ratio of 200μg/mL.

Techniques: Ex Vivo, Activation Assay, Gene Expression, Incubation, Control

Journal: Experimental physiology

Article Title: Aortic Dysfunction in Metabolic Syndrome Mediated by Perivascular Adipose Tissue TNFα and NOX2 Dependent Pathway

doi: 10.1113/EP086818

Figure Lengend Snippet: Aortic stiffness measured by A) elastic modulus in LZR and OZR rings without culture experiments (n=8, assessed by t-test). Aortic stiffness associated remodeling factors were assessed in tPVAT by B) gene expression (n=3, assessed by t-test), and C) tPVAT tissue levels of TIMP-1 (n=5, assessed by t-test. Additionally, the relative activity of MMP9 form LZR, OZR, and OZR+TNFα-AB tPVAT exudate was measured (n=5, assessed by one-ANOVA). To test the direct effect of tPVAT on aortic stiffness E) elastic modulus of donor LZR aortic rings were measured following 72-hours in culture with media, LZR-tPVAT, OZR-tPVAT, or OZR-tPVAT+TNFα-AB (n= 3–4, assessed by repeated measures ANOVA). Data expressed as Mean±SD. *denotes significant change between OZR and LZR and ^denotes significant effect of TNFα-AB treatment compared to OZR. Statistical significance was set at p<0.05. MMP, matrix metalloproteinase; TIMP, tissue inhibitor of metallo-proteinase; TNFα-AB, Tumor necrosis factor alpha neutralizing antibody.

Article Snippet: Gene Expression 50mg sections of tPVAT were incubated at 37°C in physiological HEPES buffer (43.7 mM NaCl, 80 mM KCl, 1.17 mM MgSO 4 •7H 2 O, 1.6 mM NaH 2 PO 4 , 18 mM NaHCO 3 , 0.03 mM EDTA, 5.5 mM glucose, 5 mM HEPES) or HEPES buffer containing 4μM TNFα neutralizing antibody (TNFα-AB, Catalog #: AF-510-NA, R&D systems) at a ratio of 200μg/mL.

Techniques: Gene Expression, Activity Assay