β2GPI exerts an anti-obesity effect in female mice by inhibiting lipogenesis and promoting lipolysis

Figures

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  Figure 1. Body weight of β2GPI-/- compared to WT mice recorded on a weekly basis for 16 weeks (A) Female; (B) male: ○ = NC diet WT; ● = HF diet WT, Δ = NC diet β2GPI-/-, ▲ = HF diet β2GPI-/-. Data are expressed as mean ± SD, n = 4-5 mice per group. * p < 0.05, significantly greater weight in genotype comparisons; # p <0.05, significantly greater weight in diet comparisons. NC = normal chow, HF = high fat.

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  Figure 2. VAT weight in β2GPI-/- mice fed NC or HF diet for 16 weeks (A) Female; (B) male. □ = WT, ■ = β2GPI-/-. Data are expressed as mean ± SD (n = 4-5 mice per group). *p < 0.05. NC = normal chow, HF = high fat.

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  Figure 3. Visceral adipocyte size and infiltration of VAT detected by MAC-2 staining in β2GPI-/- and WT mice fed NC or HF diet for 16 weeks Female (A, B) and male (C, D) adipocyte area and representative images of adipocyte morphology. Female (E, F) and male (G, H) crown-like structures (CLS) per high power field and representative images of macrophage infiltration of VAT. Scale bar is 50 μm. Adipocyte area is shown as mean ± SD (n = 5 mice per group, 200 cells per mouse). Number of CLS indicated by arrows in VAT was calculated as numbers per high power field in 20 high-power fields (magnification 200×). *p < 0.05. □ = WT, ■ = β2GPI-/-, NC = normal chow, HF = high fat.

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  Figure 4. Effect of β2GPI deficiency on VAT lipogenesis enzyme levels (A) Western blot analysis of VAT proteins. Quantification of the scanned images normalized to β-actin (B, D, F) and to total protein (C, E, G). Panels (B) WT mice fed a HF diet compared to WT mice fed NC. □ = NC, ■ = HF diet. ♀ = female, ♂ = male. (C) Phosphorylation of Acc (Ser79) and Atpcl (Ser 455) as a ratio of the total protein level from (B) for female mice. □ = NC diet, ■ = HF diet. (D) Female β2GPI-/- mice fed a NC diet compared to female WT mice fed a NC diet. □ = WT, ■ = β2GPI-/-. (E) p-Acc (Ser79/Acc) and p-Atpcl (Ser455). (F) Fas Protein, Acc and Atpcl. (G) Phosphorylation of Acc (Ser79) and Atpcl (Ser455) as a ratio to the total protein level from (F). □ = WT, ■ = β2GPI-/-.

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  Figure 5. Effect of β2GPI deficiency on VAT lipolysis enzyme activation (A) Western blot analysis of lipolysis enzymes in VAT of female mice. (B) Hsl – total and (C) is phosphorylation of Hsl (Ser563), Hsl (Ser660) and Hsl (Ser565). □ = NC diet, ■ = HF diet. (D) mRNA levels of Pck1, Pnpla2 and Lipe. □ = NC diet, ■ = HF diet. ♀ = female, ♂ = male. (E) Total Hsl and (F) phosphorylated Hsl (Ser563), Hsl (Ser660) and Hsl (Ser565) in female β2GPI-/- and female WT mice fed a NC diet. □ = WT, ■ = β2GPI-/-. (G) mRNA levels for Pck1 in female β2GPI-/- and female WT mice fed a NC and HF diet. □ = WT, ■ = β2GPI-/-.

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  Figure 6. Daily food intake per mouse when fed NC or HF diet for 16 weeks (A) Female; (B) male, □ = WT, ■ = β2GPI-/-. Data are expressed as mean ± SD, n = 4-5 mice per group. ***p < 0.001. NC = normal chow, HF = high fat.

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  Figure 7. Plasma leptin levels in female β2GPI-/- and WT mice fed NC or HF diet □ = WT, ■ = β2GPI-/-. Data are expressed as mean ± SD (n=5 mice per group). *p < 0.05. NC = normal chow, HF = high fat.

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  Figure 8. β2GPI deficiency and the hypothalamic appetite signaling pathways (A) Western blot analysis of hypothalamic proteins in female mice. (B-H) quantification of the immunoreactive bands (B) STAT3, (C) p-STAT3 (Tyr705) (D) SOCS3, (E) Acc, (F) p-Acc(Ser79), (G) FAS/β-actin and (H) MyD88. □ = WT, ■ = β2GPI-/-, NC = normal chow, HF = high fat. Data are expressed as mean ± SD (n = 4-5 mice per group). *p < 0.05. **p < 0.01.

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  Figure 9. Effect of a glucose tolerance challenge on plasma glucose in WT and β2GPI deficient mice fed a high fat diet (A) Female mice. (B) Male mice. Blood glucose levels were measured over 125 min after intraperitoneal injection of glucose. □ = WT, ■ = β2GPI deficient mice.