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 ³H-cortisone to ³H-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.

Key words

11β-hydroxysteroid dehydrogenase type 1 - adipose tissue biopsies - obesity - glucose

References

• 1 Tomlinson JW, Stewart PM. Cortisol metabolism and the role of 11β-hydroxysteroid dehydrogenase.  Best Pract Res Clin Endocrinol Metabol. 2001;  15 61-78
• 2 Masuzaki H, Paterson J, Shinyama H, Morton NM, Mullins JJ, Seckl JR, Flier JS. A transgenic model of visceral obesity and the metabolic syndrome.  Science. 2001;  294 2166-2170
• 3 Paterson JM, Morton NM, Fievet C, Kenyon CJ, Holmes MC, Staels B, Seckl JR, Mullins JJ. Metabolic syndrome without obesity: Hepatic overexpression of 11β-hydroxysteroid dehydrogenase type 1 in transgenic mice.  Proc Natl Acad Sci U S A. 2004;  101 7088-7093
• 4 Hermanowski-Vosatka A, Balkovec JM, Cheng K, Chen HY, Hernandez M, Koo GC, Le Grand CB, Li Z, Metzger JM, Mundt SS, Noonan H, Nunes CN, Olson SH, Pikounis B, Ren N, Robertson N, Schaeffer JM, Shah K, Springer MS, Strack AM, Strowski M, Wu K, Wu T, Xiao J, Zhang BB, Wright SD, Thieringer R. 11b-HSD1 inhibition ameliorates metabolic syndrome and prevents progression of atherosclerosis in mice.  J Exp Med. 2005;  202 517-527
• 5 Alberts P, Engblom L, Edling N, Forsgren M, Klingström G, Larsson C, Rönquist-Nii Y, Ohman B, Abrahmsén L. Selective inhibition of 11β-hydroxysteroid dehydrogenase type 1 decreases blood glucose concentrations in hyperglycaemic mice.  Diabetologia. 2002;  45 1528-1532
• 6 Wang S, Birtles S, de Schoolmeester J, Swales J, Moody G, Hislop D, O’Dowd J, Smith DM, Turnbull AV, Arch JR. Inhibition of 11β-hydroxysteroid dehydrogenase type 1 reduces food intake and weight gain but maintains energy expenditure in diet-induced obese mice.  Diabetologia. 2006;  49 1333-1337
• 7 Lee M-J, Fried SK, Mundt SS, Wang Y, Sullivan S, Stefanni A, Daugherty BL, Hermanowski-Vosatka A. Depot-specific regulation of the conversion of cortisone to cortisol in human adipose tissue.  Obesity. 2008;  16 1178-1185
• 8 Wake DJ, Homer NZM, Andrew R, Walker BR. Acute In vivo regulation of 11β-hydroxysteroid dehydrogenase type 1 activity by insulin and intralipid infusions in humans.  J Clin Endocrinol Metab 2006. 2006;  91 4682-4688
• 9 Sandeep TC, Andrew R, Homer NZM, Andrews RC, Smith K, Walker BR. Increased in vivo regeneration of cortisol in adipose tissue in human obesity and effects of the 11β-hydroxysteroid dehydrogenase type 1 inhibitor carbenoxolone.  Diabetes. 2005;  54 872-879
• 10 Rask E, Walker BR, Soderberg S, Livingstone DE, Eliasson M, Johnson O, Olsson T. Tissue-specific changes in peripheral cortisol metabolism in obese women: increased adipose 11beta-hydroxysteroid dehydrogenase type 1 activity.  J Clin Endocrinol Metab. 2002;  87 3330-3336
• 11 Wake DJ, Rask E, Livingstone DE, Soderberg S, Olsson T, Walker BR. Local and systemic impact of transcriptional up-regulation of 11β-hydroxysteroid dehydrogenase type 1 in adipose tissue in human obesity.  J Clin Endocrinol Metab. 2003;  88 3983-3988
• 12 Kannisto K, Pietilainen KH, Ehrenborg E, Rissanen A, Kaprio J, Hamsten A, Yki-Järvinen H. Overexpression of 11{beta}-hydroxysteroid dehydrogenase-1 in adipose tissue is associated with acquired obesity and features of insulin resistance: Studies in young adult monozygotic twins.  J Clin Endocrinol Metab. 2004;  89 4414-4421
• 13 Desbriere R, Vuaroqueaux V, Achard V, Boullu-Ciocca S, Labuhn M, Dutour A, Grino M. 11β-Hydroxysteroid dehydrogenase type 1 mRNA is increased in both visceral and subcutaneous adipose tissue of obese patients.  Obesity. 2006;  14 794-798
• 14 Tomlinson JW, Finney J, Gay C, Hughes BA, Hughes SV, Stewart PM. Impaired Glucose Tolerance and insulin resistance are associated with increased adipose 11β-hydroxysteroid dehydrogenase type 1 expression and elevated hepatic 5a-reductase activity.  Diabetes. 2008;  57 2652-2660
• 15 Tomlinson JW, Sinha B, Bujalska I, Hewison M, Stewart PM. Expression of 11β-hydroxysteroid dehydrogenase type 1 in adipose tissue is not increased in human obesity.  J Clin Endocrinol Metab. 2002;  87 5630-5635
• 16 Lundgren M, Svensson M, Lindmark S, Renström F, Ruge T, Eriksson J. W. Fat cell enlargement is an independent marker of insulin resistance and hyperleptinaemia.  Diabetologia. 2007;  50 625-633
• 17 Rohde JJ, Pliushchev MA, Sorensen BK, Wodka D, Shuai Q, Wang J, Fung S, Monzon KM, Chiou WJ, Pan L, Deng X, Chovan LE, Ramaiya A, Mullally M, Henry RF, Stolarik DF, Imade HM, Marsh KC, Beno DW, Fey TA, Droz BA, Brune ME, Camp HS, Sham HL, Frevert EU, Jacobson PB, Link JT. Discovery and metabolic stabilization of potent and selective 2-amino-N-(adamant-2-yl) acetamide 11β-hydroxysteroid dehydrogenase type 1 inhibitors.  J Med Chem. 2007;  50 149-164
• 18 Jang C, Obeyesekere VR, Dilley RJ, Krozowski Z, Inder WJ, Alford FP. Altered activity of 11β-hydroxysteroid dehydrogenase types 1 and 2 in skeletal muscle confers metabolic protection in subjects with type 2 diabetes.  J Clin Endocrinol Metab. 2007;  92 3314-3320
• 19 Zhu Y, Olson SH, Graham D, Patel G, Hermanowski-Vosatka A, Mundt S, Shah K, Springer M, Thieringer R, Wright S, Xiao J, Zokian H, Dragovic J, Balkovec JM. Phenylcyclobutyl triazoles as selective inhibitors of 11β-hydroxysteroid dehydrogenase type I.  Bioorg Med Chem Lett. 2008;  18 3412-3416
• 20 Lindsay RS, Wake DJ, Nair S, Bunt J, Livingstone DEW, Permana PA, Tataranni PA, Walker BR. Subcutaneous adipose 11β-hydroxysteroid dehydrogenase type 1 activity and messenger ribonucleic acid levels are associated with adiposity and insulinemia in pima Indians and Caucasians.  J Clin Endocrinol Metab. 2003;  88 2738-2744
• 21 Goedecke JH, Wake DJ, Levitt NS, Lambert EV, Collins MR, Morton NM, Andrew R, Seckl JR, Walker BR. Glucocorticoid metabolism within superficial subcutaneous rather than visceral adipose tissue is associated with features of the metabolic syndrome in South African women.  Clin Endocrinol. 2006;  65 81-87
• 22 Bujalska IJ, Kumar S, Hewison M, Stewart PM. Differentiation of adipose stromal cells: The roles of glucocorticoids and 11β-hydroxysteroid dehydrogenase.  Endocrinology. 1999;  140 3188-3196
• 23 Basu R, Singh R, Basu A, Johnson CM, Rizza RA. Effect of nutrient ingestion on total-body and splanchnic cortisol production in humans.  Diabetes. 2006;  55 667-674
• 24 Bujalska IJ, Kumar S, Stewart PM. Does central obesity reflect “Cushing's disease of the omentum”?.  Lancet. 1997;  349 (9060) 1210-1213
• 25 Hernandez-Morante JJ, Gomez-Santos C, Milagro F, Campion J, Martinez JA, Zamora S, Garaulet M. Expression of cortisol metabolism-related genes shows circadian rhythmic patterns in human adipose tissue.  Int J Obes. 2009;  33 473-480
• 26 Ryan VH, German AJ, Wood IS, Hunter L, Morris P, Trayhurn P. NGF Gene expression and secretion by canine adipocytes in primary culture: Upregulation by the inflammatory mediators LPS and TNF alpha.  Horm Metab Res. 2008;  40 861-868
• 27 Andrews RC, Herlihy O, Livingstone DEW, Andrew R, Walker BR. Abnormal cortisol metabolism and tissue sensitivity to cortisol in patients with glucose intolerance.  J Clin Endocrinol Metab. 2002;  87 5587-5589
• 28 Alberti L, Girola A, Gilardini L, Conti A, Cattaldo S, Micheletto G, Invitto C. Type 2 diabetes and metabolic syndrome are associated with increased expression of 11β-hydroxysteroid dehydrogenase 1 in obese subjects.  Int J Obes. 2007;  31 1826-1831
• 29 Meugnier E, Faraj M, Rome S, Beauregard G, Michaut A, Pelloux V, Chiasson JL, Laville M, Clement K, Vidal H, Rabasa-Lhoret R. Acute Hyperglycemia induces a global downregulation of gene expression in adipose tissue and skeletal muscle of healthy subjects.  Diabetes. 2007;  56 992-999
• 30 Walker EA, Ahmed A, Lavery GG, Tomlinson JW, Kim SY, Cooper MS, Ride JP, Hughes BA, Shackleton CH, McKiernan P, Elias E, Chou JY, Stewart PM. 11beta-Hydroxysteroid dehydrogenase type 1 regulation by intracellular glucose 6-phosphate provides evidence for a novel link between glucose metabolism and hypothalamo-pituitary-adrenal axis function.  J Biol Chem. 2007;  282 27030-27036

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