Repeated Measurements of 11β-HSD-1 Activity in Subcutaneous Adipose Tissue from Lean, Abdominally Obese, and Type 2 Diabetes Subjects – No Change Following a Mixed Meal

Abstract

The aim of this study was to measure 11β-HSD-1 activity in subcutaneous adipose tissue by an ex vivo method in three subgroups; lean, obese, and type 2 diabetes subjects, both in the fasting state and after a mixed meal and to determine the variability and reproducibility of this method. Eighteen subjects were investigated; 6 lean, 6 abdominally obese, and 6 type 2 diabetes subjects (BMI 22 ± 1, 30 ± 3 and 31 ± 3 kg/m², respectively). Needle biopsies were taken repeatedly and an index of 11β-HSD-1 activity was measured as percent conversion of (3)H-cortisone to (3)H-cortisol/100 mg tissue. For two separate biopsies taken in the fasting state on the same day, the within subjects CV was 16% and the between CV was 36% for 11β-HSD-1 activity for all subjects. For two biopsies taken in the fasting state at two different days, the total within subjects CV was 38% and the between subjects CV was 46%. Lean subjects had lower 11β-HSD-1 activity (4.8 ± 1.5% conversion of ³H-cortisone to ³H-cortisol/100 mg tissue) than both obese (14.4 ± 1.6% conversion, p<0.01) and type 2 diabetes subjects (11.7 ± 1.9% conversion, p<0.05) in the fasting state. There was no effect of a meal on 11β-HSD-1 activity in any of the three groups. The conclusions from this study are: 1) the variation coefficient for the ex vivo adipose tissue 11β-HSD-1 activity method was ∼25% for repeat measures within subjects; 2) food intake had no major impact on enzyme activity; and 3) 11β-HSD-1 activity in subcutaneous adipose tissue was significantly increased in obese subjects with or without T2DM compared to lean subjects without diabetes.

Safety, pharmacokinetics and pharmacodynamics of BI 135585, a selective 11β‐hydroxysteroid dehydrogenase‐1 (HSD1) inhibitor in humans: liver and adipose tissue 11β‐HSD1 inhibition after acute and multiple administrations over 2 weeks

“…Regulation of 11 -HSD1 activity seems to be dependent on body mass index (BMI); however, data describing the interrelationships of 11 -HSD1 expression, obesity and T2DM are conflicting. In obese subjects with and without T2DM 11 -HSD1 was found to be upregulated in subcutaneous AT compared with healthy lean controls [4,5]. In obese men with T2DM 11 -HSD1 activity in the liver did not differ from that in lean controls [6].…”

“…This hypothesis seems to be rather unlikely as INCB13739 reported >90% 11 -HSD1 enzyme inhibition in AT after a single dose in healthy and obese insulin-resistant volunteers [17] and showed efficacious glucose-and HbA1c-lowering after 12 weeks of treatment in patients with T2DM [7]. Furthermore, 11 -HSD1 seems to be similarly upregulated in the AT of healthy obese subjects and obese patients with T2DM [4]. 2.…”

Continuous inhibition of 11β‐hydroxysteroid dehydrogenase type I in adipose tissue leads to tachyphylaxis in humans and rats but not in mice

“…Ex vivo measurement of 11β-HSD1 activity in human adipose tissue Activity of human 11β-HSD1 activity in AT biopsies was measured as the conversion of 3 H-cortisone to 3 H-cortisol, following a previously reported ex vivo AT assays (Sjostrand et al, 2010). Approximately 100 mg of tissue was weighed and incubated, in triplicate, for 3 h in the presence of 3 H-cortisone (20 nM, 1 μCi · mL À1 , Amersham Biosciences GE Healthcare, Buckinghamshire, UK) containing DMEM supplemented with 6 mM glucose (Sigma Chemical Co., St Louise, MO, USA), 10% FCS (Sigma) and 1% penicillin-streptomycin (PEST; Invitrogen Corporation, Paisley, UK) in 37°C at 5% CO 2 .…”

The role and regulation of 11β-hydroxysteroid dehydrogenase type 1 in obesity and the metabolic syndrome

“…Therefore, the likely source of this increased extrasplanchnic 11β-HSD1 activity is subcutaneous adipose tissue. The presence of diabetes and/or metabolic syndrome does not appear to alter 11β-HSD1 mRNA or activity in subcutaneous adipose tissue over and above the dysregulation seen in obesity [17,[51][52][53]. Overall, these findings suggest that whole body activity in obesity-associated T2DM is increased and liver 11β-HSD1 activity is sustained, unlike in euglycaemic obesity when whole body activity is sustained in the face of reduced liver activity ( Figure 3); in both circumstances adipose 11β-HSD1 is increased.…”

“…Surprisingly, splanchnic 11β-HSD1 did not rise detectably following the meal, but because the liver accounts for at least 90% of whole body 11β-HSD1 it is difficult to imagine which other tissue could be responsible for such a large rise in whole body cortisol regeneration. In addition, subcutaneous adipose tissue 11β-HSD1 activity is not increased by a mixed meal [53]. A potential mediator of this effect is insulin, which acutely increases whole body and subcutaneous adipose tissue 11β-HSD1 activity [32].…”