Retinol binding protein 4 expression in humans: relationship to insulin resistance, inflammation, and response to pioglitazone

Abstract

Context: Retinol binding protein 4 (RBP4) was recently found to be expressed and secreted by adipose tissue, and was strongly associated with insulin resistance.

Objective: The aim was to determine the relationship between RBP4 and obesity, insulin resistance, and other markers of insulin resistance in humans.

Design and patients: RBP4 mRNA levels in adipose tissue and muscle of nondiabetic human subjects with either normal or impaired glucose tolerance (IGT) were studied, along with plasma RBP4. RBP4 gene expression was also measured in adipose tissue fractions, and from visceral and sc adipose tissue (SAT) from surgical patients.

Setting: The study was conducted at University Hospital and General Clinical Research Center.

Intervention: Insulin sensitivity (S(I)) was measured, and fat and muscle biopsies were performed. In IGT subjects, these procedures were performed before and after treatment with metformin or pioglitazone.

Main outcome measures: The relationship between RBP4 expression and obesity, S(I), adipose tissue inflammation, and intramyocellular lipid level, and response to insulin sensitizers was measured.

Results: RBP4 was expressed predominantly from the adipocyte fraction of SAT. Although SAT RBP4 expression and the plasma RBP4 level demonstrated no significant relationship with body mass index or S(I), there was a strong positive correlation between RBP4 mRNA and adipose inflammation (monocyte chemoattractant protein-1 and CD68), and glucose transporter 4 mRNA. Treatment of IGT subjects with pioglitazone resulted in an increase in S(I) and an increase in RBP4 gene expression in both adipose tissue and muscle, but not in plasma RBP4 level, and the in vitro treatment of cultured adipocytes with pioglitazone yielded a similar increase in RBP4 mRNA.

Conclusions: RBP4 gene expression in humans is associated with inflammatory markers, but not with insulin resistance. The increase in RBP4 mRNA after pioglitazone treatment is unusual, suggesting a complex regulation of this novel adipokine.

FIG. 1

Relationships between plasma RBP4 and BMI (A) and S_I (B–D). Plasma RBP4 was measured with either ELISA assay (A and B) or Western blotting assay (C and D), as described in Subjects and Methods. The data in D panel were generated from the mean of four repeated Western blots. AU, Arbitrary units.

FIG. 2

Relationship between adipose tissue GLUT4 mRNA and RBP4 expression (A) and plasma RBP4 (B). Relationship between adipose tissue RBP4 and CD68 (C) and MCP1 (D). Levels of RBP4 mRNA, GLUT4 mRNA, MCP1 mRNA, and CD68 mRNA were determined by real-time RT-PCR as described in Subjects and Methods. Levels of plasma RBP4 were determined by ELISA assay.

FIG. 3

Effects of pioglitazone and metformin treatment on RBP4 expression in adipose tissue (A), muscle tissue (B), differentiated SGBS adipocytes (C), and on plasma RBP4 level (D). *, P = 0.001 vs. baseline of adipose tissue. †, P = 0.02 vs. baseline of muscle tissue. ¥, P < 0.05 vs. baseline of SGBS cells. Post Tx, After treatment.