Deletion of Seipin Attenuates Vascular Function and the Anticontractile Effect of Perivascular Adipose Tissue

Figures

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  Figure 1. Insulin resistance in WT and Seipin deleted mice. (A) Plasma triglyceride (TG), total cholesterol (TC), glucose content in mice fasted for 4 h. (B) Free fatty acid (FFA) in mice fasted for 4 h. (C) Plasma levels of IL-6, TNF-α in mice fasted for 4 h. (D) Plasma insulin, leptin, adiponectin content in mice fasted for 4 h. (E) Glucose tolerance tests and insulin tolerance tests (F) performed on 6-month-old Seipin−/− and WT mice. Data are expressed as mean ± SEM. N = 6. *P < 0.05, **P < 0.01 for Seipin−/− mice vs. WT.

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  Figure 2. Reduced PVAT mass in Seipin deleted mice. (A) Detection of Seipin mRNA and protein expression in various tissues of WT mice. N = 6. (B) Body weight and (C) Mass of fat pad and PVAT in 6-month-old Seipin−/− and WT mice. Brown adipose tissue (BAT), White adipose tissue (WAT), aortic perivascular adipose tissue (Ao PVAT), mesenteric PVAT (Mes PVAT). Data are expressed as mean ± SEM. N = 6. *P < 0.05, **P < 0.01 for Seipin−/− mice vs. WT.

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  Figure 3. Histological observation of PVAT from WT and Seipin−/− mice. (A) Representative H&E and sirius-red staining of PVAT from 6-month-old WT and Seipin−/− mice. (B) Representative electron microscopy of PVAT from 6-month-old WT and Seipin−/− mice. (C) Mac2 immunostaining of PVAT from 6-month-old WT and Seipin−/− mice. m, mitochondria; n, adipocyte nucleus; L, lipid droplets. Pink arrow indicated deformed mitochondria.

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  Figure 4. Inflammation, fibrosis and ER stress gene expression in PVAT and aorta from WT and Seipin−/− mice. Relative mRNA levels of macrophages and inflammatory cytokines (A,B), fibrosis (C), and ER stress (D) in PVAT and aorta from 6-month-old WT and Seipin−/− mice. N = 6. (E,F) Western blot images and densitometric quantitation for the indicated proteins and phosphoproteins related to inflammation and ER stress in PVAT from WT and Seipin−/− mice. Data are expressed as mean ± SEM. N = 6, *P < 0.05, **P < 0.01 for Seipin−/− mice vs. WT.

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  Figure 5. Vascular lesion in aorta of Seipin−/− mice. (A) Representative H&E and Gomrori's aldehyde-fuchin staining in thoracic aorta, abdominal aorta and mesenteric artery in WT and Seipin−/− mice. (B) Electron microscopy of thoracic aorta from 6-month-old Seipin−/− and WT mice. Red arrow indicated derangement of elastic fibers in aorta. Thin red arrow showed deformed mitochondria. Green arrow indicated dilated ER. m, mitochondria; ER, endoplasmic reticulum.

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  Figure 6. Impaired vasoactivity in Seipin−/− mice. Systolic (A) and diastolic (B) of WT and Seipin−/− mice. N = 10. (C) Phenylephrine induced contraction of aorta rings with (+) or without (-) PVAT prepared from 6-month-old WT and Seipin−/− mice. (D, E, F) Phenylephrine dose-response curves of mesenteric artery rings with (+) or without (-) PVAT prepared from 6-month-old WT and Seipin−/− mice. (G, H) Endothelial-dependent relaxation with acetylcholine (Ach) and smooth muscle relaxation with sodium nitroprusside (SNP) in aortic rings prepared from 6-month-old WT and Seipin−/− mice. N = 10. (I) Representative curves of transferring 5 mL incubation of aorta PVAT of WT and Seipin−/− mice to aorta without PVAT after preconstruction with phenylephrine (PHE, 10 uM). *P < 0.05, **P < 0.01 for Seipin−/− mice vs. WT.