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. 2008 Oct;295(4):E842-50.
doi: 10.1152/ajpendo.90359.2008. Epub 2008 Jul 29.

Adiponectin secretion and response to pioglitazone is depot dependent in cultured human adipose tissue

Susan A Phillips  1 , Theodore P Ciaraldi, Deborah K Oh, Michelle K Savu, Robert R Henry
Affiliations

Adiponectin secretion and response to pioglitazone is depot dependent in cultured human adipose tissue

Susan A Phillips et al. Am J Physiol Endocrinol Metab. 2008 Oct.

Abstract

The subcutaneous (S) and visceral (V) adipose tissue (AT) depots are increasingly recognized as distinct. To test the hypothesis that depot differences exist for adiponectin, fresh and cultured human VAT and SAT from obese type 2 diabetic (T2D) and obese nondiabetic (ND) subjects was examined to determine whether differences in adiponectin content and secretion occurred as a function of depot studied, diabetic status, and response to thiazolidinedione treatment. VAT and SAT were obtained by biopsy and AT explants cultured in defined media for 7 days. Protein expression was assessed by Western blot. Adiponectin content of conditioned medium was determined by radioimmunoassay. Diabetic status had no effect on adiponectin secretion over days 0-2 of culture. In ND SAT, secretion fell over days 2-4 but was sustained at greater levels vs. T2D SAT. In both ND and T2D VAT, adiponectin secretion was low, similar to T2D SAT. Over the 7-day culture period, cellular adiponectin increased in ND SAT and VAT; it remained unchanged in T2D SAT and VAT. Pioglitazone increased adiponectin secretion and content in all SAT. Pioglitazone failed to increase adiponectin secretion from either ND or T2D VAT and increased cellular content only in ND VAT. AT depot differences exist in the secretion of adiponectin and responsiveness to thiazolidinedione treatment. These data suggest that SAT, not VAT, appears to be the major contributor to increased circulating adiponectin levels in response to pioglitazone treatment.

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Figures

Fig. 1.
Fig. 1.
Adiponectin secretion by nondiabetic (ND; A) and type 2 diabetic (T2D; B) adipose tissue (AT) explants. Explants were cultured for 7 days in F-10 defined medium (DM) containing 30 nM Dex and 1 nM insulin. Medium was sampled every 48 h and analyzed by RIA for adiponectin (Ad) content. Results are reported as average ± SE; n = 10 for ND subcutaneous (S)AT and n = 8 for ND visceral (V)AT; n = 13 for T2D SAT and n = 8 for T2D VAT. Open bars, SAT; filled bars, VAT. *P < 0.05 vs. secretion on day 2.
Fig. 2.
Fig. 2.
Cell content of adiponectin in adipocytes from freshly isolated and cultured AT. Adipocyte protein extracts were prepared from fresh (F) and 7-day-cultured (Cx) ND and T2D AT and analyzed by immunoblotting with anti-adiponectin antibody as described in materials and methods. Top: representative autoradiographs. Bottom: quantitation of autoradiographs. Adiponectin expression is presented as a percentage of the value determined from analysis of fresh cells from the same individual, average ± SE; n = 9 for ND and n = 7 for T2D SAT; n = 12 for ND and n = 8 for T2D VAT. Open bars, ND; hatched bars, T2D. *P < 0.05 vs. value in freshly isolated adipocytes.
Fig. 3.
Fig. 3.
Adiponectin secretion by cultured ND SAT (A) and VAT (B) explants: effect of PPARγ agonist pioglitazone (Pio) treatment. Explants were cultured for 7 days in DM ± Pio (10 μM). Medium was sampled every 2 days and analyzed by RIA for adiponectin content. Results are reported as average ± SE; n = 12 for SAT and n = 9 for VAT. open bars, control; hatched bars, +Pio. *P < 0.05 vs. secretion on day 2; †P < 0.05 vs. paired control.
Fig. 4.
Fig. 4.
Adiponectin secretion by cultured T2D SAT (A) and VAT (B) explants: effect of PPARγ agonist Pio treatment. Explants were cultured for 7 days in DM ± Pio (10 μM). Medium was sampled every 2 days and analyzed by RIA for adiponectin content. Results are reported as average ± SE; n = 13 for SAT and n = 10 for VAT. Open bars, control; hatched bars, +Pio. *P < 0.05 vs. secretion on day 2; †P < 0.05 vs. paired control.
Fig. 5.
Fig. 5.
Leptin secretion by cultured SAT and VAT explants: effect of PPARγ agonist Pio treatment. Leptin secretion by SAT (A) and VAT (B) from combined ND and T2D AT explants. Explants were cultured for 7 days in DM containing 30 nM Dex and 1 nM insulin ± 10 μM Pio. Medium was sampled every 2 days and analyzed by RIA for leptin content. Results are reported as average ± SE. SAT: n = 16 −Pio and n = 8 +Pio; VAT: n = 14 −Pio and n = 10 +Pio. VAT. Open bars, control; hatched bars, +Pio. *P < 0.05 vs. secretion on day 2; †P < 0.05 vs. paired control.
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