Postprandial fatty acid uptake and adipocyte remodeling in angiotensin type 2 receptor-deficient mice fed a high-fat/high-fructose diet

Figures & data

Table 1. Anthropometric and biochemical markers.

	WT SD	WT HFHF-STZ	AT2R-KO SD	AT2R-KO HFHF-STZ
Weight gain (%)	19.4 ± 2.2	21.7 ± 2.5	5.9 ± 1.5^a	10.9 ± 2.7^b
Blood glucose (mM)	4.2 ± 0.2	6.1 ± 0.5^a	7.3 ± 0.5^a	8.7 ± 0.7^b
Blood glucose AUCOGTT (mM.120 min^−1)	2131 ± 98	2155 ± 84	2029 ± 111	2248 ± 69
Plasma NEFA (µM)	604 ± 87	558 ± 74	708 ± 40	667 ± 70
Plasma triglycerides (µM)	371 ± 33	286 ± 41	301 ± 33	264 ± 46
Plasma insulin (pg.mL^−1)	1993 ± 726	1040 ± 329	309 ± 110^a	293 ± 179
Plasma leptin (pg.mL^−1)	1718 ± 394	2731 ± 899	715 ± 207^a	1821 ± 737
Heart weight (mg)	158.4 ± 5.6	134.2 ± 2.4^a	129.0 ± 5.2^a	135.8 ± 7.0
Liver weight (mg)	1233 ± 47	906 ± 43^a	1152 ± 76	961 ± 8^c
Skeletal muscle (Gastrocnemius) weight (mg)	117.6 ± 10.2	125.5 ± 15.0	105.8 ± 8.4	125.0 ± 9.9
Visceral adipose tissue weight (mg)	903.1 ± 125.6	771.8 ± 162.0	648.2 ± 152.6	878.0 ± 193.2

Values are means ± SE from 9–10 animals per group. Wild-type (WT) and AT2R-knockout-out (KO) mice were fed for 6 weeks either with standard laboratory rodent diet (SD) or with a high-fat/high-fructose with small injection of streptozotocin (HFHF-STZ). Statistical analysis was performed using Mann-Whitney test. a : P < 0 .05 vs. WT SD ; b : P < 0 .05 vs. WT HFHF-STZ ; c : P < 0 .05 vs. AT2R-KO SD.

Table 2. Food and water consumption per day.

	WT SD	WT HFHF-STZ	AT2R-KO SD	AT2R-KO HFHF-STZ
Mean water consumption (mL.day^−1)	6.1 ± 0.2	4.8 ± 0.1^a	5.4 ± 0.1^a	5.1 ± 0.1
Mean food consumption (g.day^−1)	4.5 ± 0.2	2.3 ± 0.1^a	3.7 ± 0.1^a	2.3 ± 0.1^c
Mean caloric intake (kcal.day^−1)	15.3 ± 0.8	11.2 ± 0.4^a	12.7 ± 0.2^a	11.6 ± 0.5^c

Values are means ± SE from 9–10 animals per group. Wild-type (WT) and AT2R-knockout-out (KO) mice were fed for 6 weeks either with standard laboratory rodent diet (SD) or with a high-fat/high-fructose with small injection of streptozotocin (HFHF-STZ). Statistical analyses were performed using Mann-Whitney test. a: P < 0 .05 vs. WT SD; b: P < 0 .05 vs. WT HFHF-STZ ; c: P < 0 .05 vs. AT2R-KO SD.

Figure 1. Effect of HFHF-STZ on non-esterified fatty acid uptake of [¹⁸F]-FTHA in WT and AT2R-KO mice. WT and AT2R-KO mice were fed either a standard laboratory rodent diet (SD) or a high-fat/high-fructose diet with small injection of streptozotocin (HFHF-STZ) for 6 weeks. At the end of the experimental period, [¹⁸F]-FTHA was given i.v. during the fasting state. The K_i fractional uptake of [¹⁸F]-FTHA was analyzed by µPET in the heart (A) and the liver (C); and the K_m net uptake of [¹⁸F]-FTHA was analyzed by µPET in the heart (B) and the liver (D). Data are presented as mean ± SE (n = 8–10). Statistical analyses of the data were performed using Mann-Whitney test.

Figure 2. Effect of HFHF-STZ on postprandial dietary fatty acid uptake of [¹⁸F]-FTHA in WT and AT2R-KO mice. WT and AT2R-KO mice were fed either a standard laboratory rodent diet (SD) or a high-fat/high-fructose diet with small injection of streptozotocin (HFHF-STZ) for 6 weeks. At the end of the experimental period, [¹⁸F]-FTHA was given per os during the postprandial state. The uptake of [¹⁸F]-FTHA was analyzed in the heart (A), the liver (B), the skeletal muscle (gastrocnemius) (C), the kidney (D), the visceral (retroperitoneal) adipose tissue (E) and the subcutaneous adipose tissue (F). Data are presented as mean ± SE (n = 8–10). Statistical analyses of the data were performed using Mann-Whitney test.

Figure 3. Effect of HFHF-STZ on adipocyte size distribution in WT and AT2R-KO mice. Mice were fed a standard diet (SD) or high-fat/high-fructose diet with STZ (HFHF-STZ) for 6 weeks. Adipocyte size distribution (A, B) and areas (C, D) from subcutaneous adipose tissue (A, C) and visceral (retroperitoneal) (B, D) adipose tissue. Data are presented as mean ± SE (n = 8–10). Statistical analyses were performed using one-way ANOVA followed by the Tukey's multiple comparisons test. (A, C), Statistical significance.

Figure 4. H&E staining of adipose tissue from WT and AT2R-KO mice, after 6 weeks fed a standard diet (SD) or high-fat/high-fructose diet with STZ (HFHF-STZ). Sections (5 μm) of subcutaneous (A-D) and visceral (retroperitoneal) adipose tissues (E-H) were stained with H&E. Ten images per histological section were used for analysis. Images were acquired using a Leica microscope equipped with a 10X objective. Scale bar, 40 μm.

Figure 5. µTEP protocol. Timeline used to observe intravenous uptake of [¹⁸F]-FTHA in mice (A) and an example of a coronal slice through the µPET image of FTHA uptake in a WT mouse on a normal diet obtained by this protocol (B). Dyn: Dynamic PET.