Decrease of Perivascular Adipose Tissue Browning Is Associated With Vascular Dysfunction in Spontaneous Hypertensive Rats During Aging

Figures

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  FIGURE 1. Characteristic differences among sWAT, iBAT, and tPVAT. (A) Representative HE and UCP1 staining images of sWAT, iBAT, and tPVAT from 8-week-old WKY rats. Magnification bar = 100 μm. (B) Quantitative analysis of cell numbers and UCP1 signals in sWAT, iBAT, and tPVAT from WKY rats (n = 4). (C) Western blot analysis of UCP1, PGC1α, and PPARγ protein levels in sWAT, iBAT, and tPVAT. GAPDH serves as internal control. Quantitative results are shown on the right (n = 4). ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.

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  FIGURE 2. Browning level of tPVAT increases in SHR during aging. (A) The body weight of 8-week-old and 16-week-old WKY and SHR (n = 10). (B) The SBP and DBP of 8-week-old and 16-week-old WKY and SHR (n = 10). (C) Representative HE and UCP1 staining images of the tPVAT of 8-week-old and 16-week-old WKY and SHR. Magnification bar = 50 μm. (D) Quantitative analysis of cell numbers and UCP1 signals in tPVAT from 8-week-old and 16-week-old WKY and SHR rats (n = 4). (E) Western blot analysis of UCP1, PGC1a, and PPARγ expression levels in the tPVAT of 8-week-old and 16-week-old WKY and SHR. Quantitative results are shown on the right (n = 4). ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.

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  FIGURE 3. Thoracic aorta reactivity incubated with tPVAT from WKY or SHR. (A) Illustration of bioassay protocol. (B) Average percent relaxation of thoracic aorta to ACH with or without 12 h tPVAT treated (n = 6). (C) Representative raw tracing of diastolic response to ACH after PHE (3 μM) pre-contraction. The thoracic aorta rings were collected from 8-week-old WKY rats and the PVAT adjacent to the rings was cleaned. The donor tPVAT were collected from 8-week-old and 16-week-old WKY or SHR. The rings were incubated with tPVAT in DMEM for 12 h at 37°C. (D) Average percent relaxation to ACH of thoracic aorta treated with 8-week-old or 16-week-old WKY or SHR tPVAT (n = 6). (E) Average percent relaxation to ACH of thoracic aorta treated with 8-week-old or 16-week-old WKY or SHR tPVAT (n = 6). (F) Representative raw traces showing dose-dependent aortic ring constriction in response to PHE after KCl (60 mM) pre-contraction. (G) Average percent contraction to PHE of thoracic aorta treated with 8-week-old or 16-week-old WKY or SHR tPVAT (n = 6). ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.

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  FIGURE 4. CD73 expression and adenosine production in the tPVAT of SHR decrease during aging. (A) Western blot analysis of CD73 protein level in the tPVAT of 8-week-old and 16-week-old WKY and SHR. Quantitative results are shown on the lower panel (n = 4). (B) Adenosine concentration (ELISA) in the tPVAT-conditional media from 8-week-old and 16-week-old WKY and SHR (n = 4). ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.