MiR-125a expression in adipose tissue of obese patients and correlations with physiological parameters.

<p>MiR-125a expression was quantified by qRT-PCR in subcutaneous (A) and visceral adipose tissue (B) in obese patients (Ob) with or without diabetes (Diab) and controls. MiR-125a expression level in obese patients was correlated with total triglycerides level, circulating leptin, γ-glutamyltransferase (γ-GT) and adiponectin levels, fat mass and fat free mass percentages (C) in males (▪) and females (•). Expression data were normalized with miR-16, an endogenous miR whom expression level do not vary across group of study. Data are the mean ± SEM of 6 non obese subjects for subcutaneous adipose tissue, 7 non obese subjects for visceral adipose tissue, 9 obese non diabetic and 7 obese diabetic patients. Parametric Pearson correlation tests were performed. *p<0.05, **p<0.01, ***p<0.001 significant differences between obese patients and controls.</p

MiR-125a target gene expression regulation in adipose tissue of obese patients.

<p>Expression of <i>Tef</i>, <i>Masp1</i>, <i>Rtn2</i>, <i>Ube2l3</i> and <i>Adam9</i> was quantified by qRT-PCR in subcutaneous (A) and visceral (B) adipose tissue of obese patients (Ob) with diabetes (Diab) or without (NonDiab) and non obese subjects (Controls). Expression levels were normalized with human <i>β-Actin</i>. ^$p<0.05, ^$$p<0.01, ^$$$p<0.001 significantly different to non obese subjects.</p

Phenotype characterization of the effects of 1 and 15 weeks of HFD feeding in C57BL/6J and BALB/c mice (CHD, Carbohydrate diet; HFD, High-fat diet).

<p>*p<0.05, **p<0.01, ***p<0.001 significant differences between mice fed with HFD and CHD.</p

Biological relationships between insulin resistance and miR-125a expression.

<p>MiR-125a expression levels were quantified by q (quantitative) RT-PCR in adipose tissue of mice fed HFD for 1 week (A) or 15 weeks (B). The effect of <i>in vitro</i> insulin resistance (C) on miR-125a expression (D) was tested by qRT-PCR in 3T3-L1 adipocytes. Expression of miR-125a was normalized with small nucleolar (sno) RNA202 in adipose tissue and U6 in cultured cells. Data are expressed as mean ± SEM for 4–5 mice per group and for 3 replicated experiments in cultured cells. *p<0.05 significant differential expression of miR-125a between HFD-fed and control mice of the same strain. ^£££p<0.001 significant differences in glucose transport stimulated by insulin in 3T3-L1 adipocytes. ∧∧p<0.01 significant differences in miR-125a expression between IR-3T3-L1 adipocytes and control-3T3-L1 adipocytes.</p

Patient physiological parameters (F, Female; M, Male; BMI, Body mass index; TGL, Triglyceride; γGT, γ-glutamyltransferase; IL6, Interleukin 6; ApoB, Apolipoprotein B;QUICKI, Quantitative insulin sensitivity index).

<p>Patient physiological parameters (F, Female; M, Male; BMI, Body mass index; TGL, Triglyceride; γGT, γ-glutamyltransferase; IL6, Interleukin 6; ApoB, Apolipoprotein B;QUICKI, Quantitative insulin sensitivity index).</p

Expression patterns of miR-125a target gene sets in fat fed mice and in insulin resistant adipocytes.

<p>Gene set enrichment profiles of miR-125a gene targets (A) were determined for BALB/c mice after 1 week of HFD feeding (left) and C57BL/6J mice after 15 weeks of HFD feeding (right). Heatmap of miR-125a leading edge subsets are shown for fat fed C57BL/6J (top) and BALB/c (bottom) mice (B), alongwith enriched target genes significantly differentially expressed in these mice (C). Expression of <i>Tef</i>, <i>Masp1</i>, <i>Rtn2</i>, <i>Ube2l3</i> and <i>Adam9</i> was quantified by qRT-PCR in IR-3T3-L1 (▪) and control (□) adipocytes (D). Expression level was normalized with mouse <i>β-Actin</i>. Data are the mean ± SEM of 4 cultured cell replicates. *p<0.05 significant differences between IR-3T3-L1 adipocytes and control-3T3-L1 adipocytes.</p

Circulating inflammatory markers

<p>VAL treatment (<i>n</i> = 17) did not significantly affect plasma concentrations of (<b>A</b>) MCP-1, (<b>B</b>) TNF-α, (<b>C</b>) adiponectin and (<b>D</b>) leptin compared with PLB (<i>n</i> = 19). MCP-1, monocyte chemoattractant protein-1; TNF-α, tumour necrosis factor-α.</p

Correlation between the change in adipocyte diameter and insulin sensitivity after 26-wks VAL or PLB treatment (<i>n</i> = 30)

<p>The decrease in adipocyte size was significantly associated with alterations in insulin sensitivity after VAL (n = 14, closed circles) and PLB (n = 16, open circles) treatment (<i>r</i> = −0.452, <i>P</i> = 0.012).</p

Enrollment flow chart.

<p>Enrollment flow chart.</p

Subject characteristics before and after 26-wks treatment with VAL or PLB.

<p>BMI, body mass index; HbA1C, glycated haemoglobin; NEFA, non-esterified fatty acid; SBP, systolic blood pressure; DBP, diastolic blood pressure; TAG, triacylglycerol; WHR, waist-to-hip ratio; <i>N.S.</i>, not significant. ^*VAL <i>vs.</i> PLB treatment assessed by repeated-measures ANOVA. ^#<i>P</i><0.05 <i>vs</i>. PLB. Values are means±SEM.</p