Association of rs939347 with obesity features.

<p>A) Waist circumference in men. B) BMI in men C) Waist circumference in women D) BMI in women.</p

Association between rs939347 and the risk of obesity by gender.

<p>Obese is defined as having a BMI>30 kg/m². Lean is defined as having a BMI<26 kg/m². Abbreviations: CI, confidence interval; OR, odds ratio.</p

<i>REV-ERB ALPHA</i> Polymorphism Is Associated with Obesity in the Spanish Obese Male Population

<div><p><i>REV-ERB ALPHA</i> has been shown to link metabolism with circadian rhythms. We aimed to identify new polymorphisms in the promoter of <i>REV-ERB ALPHA</i> and tested whether these polymorphisms could be associated with obesity in the Spanish population. Of the 1197 subjects included in our study, 779 were obese (BMI 34.38±3.1 kg/m²) and 418 lean (BMI 23.27±1.5 kg/m²). In the obese group, 469 of the 779 had type 2 diabetes. Genomic DNA from all the subjects was obtained from peripheral blood cells and the genotyping in the <i>REV-ERB ALPHA</i> promoter was analyzed by High Resolution Melting. We found six polymorphisms in the <i>REV-ERB ALPHA</i> promoter and identified rs939347 as a SNP with the highest frequency in the total population. We did not find any association between rs939347 and type 2 diabetes (p = 0.101), but rs939347 was associated with obesity (p = 0.036) with the genotype AA exhibiting higher frequency in the obese (5.2% in total obese vs 2.4% in lean). This association was found only in men (p = 0.031; 6.5% AA-carriers in obese men vs 1.9% AA-carriers in lean men), with no association found in the female population (p = 0.505; 4.4% AA-carriers in obese women vs 2.7% AA-carriers in lean women). Our results suggest that the <i>REV-ERB ALPHA</i> rs939347 polymorphism could modulate body fat mass in men. The present work supports the role of <i>REV-ERB ALPHA</i> in the development of obesity as well as a potential target for the treatment of obesity.</p></div

Association of risk genotype AA vs AG+GG in rs939347 with study variables.

<p>Results of logistic regresion models of having the genotype AA versus having the genotype AG and/or the genotype GG in the SNP 939347 by increasing levels of the aforementioned variables. *Multivariate models were creating adjusting by age and gender. **if waist circumference >94 cm in men and >80 cm in women, according to EGIR criteria (1999).</p

Association between rs939347 and the risk of obesity.

<p>Obesity risk is defined as BMI > 30 kg/m². Abbreviations: CI, confidence interval; OR, odds ratio; SNP, single-nucleotide polymorphism. <i>P</i> values were adjusted for age and gender.</p

Genotype distribution of rs939347 among lean, obese and T2D subjects by gender.

<p>P1 =  Lean vs total obese population; P2 =  Controls vs Obese (OB); P3 =  Controls vs type 2 diabetes (T2D); P4 =  Ob vs T2D.</p

Clinical characteristics of the subjects.

<p>P1 =  Lean vs total obese population; P2 =  Lean vs Obese (OB); P3 =  Lean vs type 2 diabetes (T2D); P4 =  OB vs T2D.</p

β-cell mass recovery after streptozotocin-induced diabetes in PTP1B ⁻/⁻ mice.

<p>A) Blood glucose levels along the experimental period (7 weeks) after STZ-induced β-cell damage in PTP1B ⁻/⁻ and WT fed mice. B) Body weight at the end of the experimental period after STZ-induced β-cell damage in PTP1B ⁻/⁻ and WT fed mice. C) Body weight gain 7 weeks after STZ injection in both experimental groups. D) Pancreas weight (normalized by body weight) of STZ-treated PTP1B ⁻/⁻ and WT fed mice 7 weeks after STZ injection. E) β-cell mass is quantified blindly as β-cell volume density, multiplied by pancreas weight (n = 6 animals per group). Representative images showing immunostaining for insulin (red), glucagon (green), and Dapi for nuclei (blue) on pancreatic sections from STZ-treated PTP1B ⁻/⁻ and WT mice. F) Levels of proliferating β-cells (ki67+/insulin+) in STZ-treated PTP1B ⁻/⁻ and WT mice (n = 6 animals per group). Representative images showing immunostaining for insulin (red), ki67 (green), and Dapi for nuclei (blue) on pancreatic sections from STZ-treated PTP1B ⁻/⁻ and WT mice. G) Levels of apoptotic β-cells (caspase3+/insulin+) in STZ-treated PTP1B ⁻/⁻ and WT mice. (n = 6 animals per group). Representative images showing immunostaining for insulin (red), Caspase3 (green), and Dapi for nuclei (blue) on pancreatic sections from STZ-treated PTP1B ⁻/⁻ and WT mice. All results represent mean±SEM; * p<0.05, ** p<0.005 PTP1B ⁻/⁻ + STZ <i>vs</i> WT + STZ.</p

Glucose induced insulin secretion both <i>in vivo</i> and in isolated islets from PTP1B ⁻/⁻ and WT mice.

<p>Islets were isolated from 8 weeks old WT and PTP1B ⁻/⁻ mice and insulin secretion was assayed at indicated glucose concentrations (2.8 and 16.7 mM) in static incubation experiments as indicated in materials and methods section. A) Insulin secretion per islet. B) Islet insulin content. C) Insulin secretion as a percentage of total insulin content. D) Plasma insulin levels after glucose administration during an ipGTT test in PTP1B ⁻/⁻ (n = 5) and WT (n = 5) mice. E) Glucose tolerance test (ipGTT): blood glucose levels of PTP1B ⁻/⁻ (n = 13) and WT (n = 12) mice at the indicated time points after an intraperitoneal injection of glucose (2 g/Kg body weight). F) ipGTT area under the curve (AUC). AUC was calculated using the trapezoidal rule. All results represent mean±SEM * p<0.05; ** p<0.005 PTP1B ⁻/⁻<i>vs</i> WT.</p

Ablation of PTP1B increases proliferation in <i>in vitro</i> transfected MIN6 cells and dispersed islet cells.

<p>A) mRNA expression and B) protein content are used to confirm the efficiency of PTP1B siRNA transfection and silencing in MIN6 cells. Values are expressed relative to 36B4 as a housekeeping gene and actin protein content respectively. C) <i>In vitro</i> BrdU incorporation in proliferating si-ptpn1 MIN6 cells. Represented values are normalized to scrambled siRNA (Sc) MIN6 cells; n = 3 different experiments, 10 replicates per group each experiment. D) <i>In vitro</i> BrdU incorporation in PTP1B ⁻/⁻ proliferating dispersed islet cells is expressed relative to WT; n = 3 different experiments, 6 replicates per group each experiment. All bars represent mean±SEM * p<0.05, ** p<0.005 PTP1B ⁻/⁻<i>vs</i> WT.</p