Distinguishing the Activities of 11beta-Hydroxysteroid Dehydrogenases in Vivo Using Isotopically Labeled Cortisol

@article{citeulike:1324783, abstract = {The isozymes of 11{beta}-hydroxysteroid dehydrogenase (11{beta}HSDs) catalyze the interconversion of cortisol and cortisone. The type 2 dehydrogenase inactivates cortisol to cortisone, whereas the type 1 catalyzes predominantly the reverse reductive reaction. These reactions take place in different tissues, where they are subject to distinct regulation, and may be important in common pathologies. Current methods to determine the activities of these enzymes in vivo rely only on the balance between cortisol and cortisone, do not measure turnover, and cannot distinguish between the two reactions. We have investigated the use of [9,11,12,12-2H4]cortisol (d4F) to distinguish the dehydrogenase and reductase activities. On metabolism by dehydrogenation, d4F loses 11{alpha}- deuterium, forming trideuterated cortisone (d3E) and is regenerated by reduction to trideuterated cortisol (d3F). Healthy men (n = 6) participated in a randomized, double blind, cross-over study comparing oral placebo and the 11{beta}HSD inhibitor, carbenoxolone (100 mg every 8 h for 7 d). d4F and its metabolites were measured in plasma and urine during a steady state infusion. Inhibition of 11{beta}HSDs by carbenoxolone was measured by increased steady state concentrations of d4F (41 {+/-} 5.1 vs. 48 {+/-} 7.7 nM; P < 0.05) and a fall in the rate of appearance of d3F (P < 0.05). 11{beta}HSD1 reductase activity could be measured specifically as conversion of d3E to d3F (28 {+/-} 4.2 vs. 17 {+/-} 3.1 nM; P < 0.05), whereas 11{beta}HSD2 could be measured by initial rates of appearance of d3E or from urinary ratios of d4F/(d3E + d3F) (0.73 {+/-} 0.06 vs. 1.02 {+/-} 0.03; P < 0.05). This technique offers a significant advance in the methods available to measure turnover in 11{beta}HSDs and isozymes of 11{beta}HSDs in vivo in human studies, and this study confirms that carbenoxolone inhibits both isozymes of 11{beta}HSD. 10.1210/jc.87.1.277}, author = {Andrew, Ruth and Smith, Kenneth and Jones, Gregory C. and Walker, Brian R. }, citeulike-article-id = {1324783}, doi = {http://dx.doi.org/10.1210/jc.87.1.277}, journal = {J Clin Endocrinol Metab}, keywords = {11bhsd1, 11bhsd2, cortisol, isotope, tracer}, month = {January}, number = {1}, pages = {277--285}, posted-at = {2007-05-24 11:29:44}, priority = {2}, title = {Distinguishing the Activities of 11{beta}-Hydroxysteroid Dehydrogenases in Vivo Using Isotopically Labeled Cortisol}, url = {http://dx.doi.org/10.1210/jc.87.1.277}, volume = {87}, year = {2002} }

TY - JOUR ID - citeulike:1324783 L3 - citeulike-article-id:1324783 TI - Distinguishing the Activities of 11{beta}-Hydroxysteroid Dehydrogenases in Vivo Using Isotopically Labeled Cortisol JF - J Clin Endocrinol Metab VL - 87 IS - 1 SP - 277 EP - 285 N2 - The isozymes of 11{beta}-hydroxysteroid dehydrogenase (11{beta}HSDs) catalyze the interconversion of cortisol and cortisone. The type 2 dehydrogenase inactivates cortisol to cortisone, whereas the type 1 catalyzes predominantly the reverse reductive reaction. These reactions take place in different tissues, where they are subject to distinct regulation, and may be important in common pathologies. Current methods to determine the activities of these enzymes in vivo rely only on the balance between cortisol and cortisone, do not measure turnover, and cannot distinguish between the two reactions. We have investigated the use of [9,11,12,12-2H4]cortisol (d4F) to distinguish the dehydrogenase and reductase activities. On metabolism by dehydrogenation, d4F loses 11{alpha}- deuterium, forming trideuterated cortisone (d3E) and is regenerated by reduction to trideuterated cortisol (d3F). Healthy men (n = 6) participated in a randomized, double blind, cross-over study comparing oral placebo and the 11{beta}HSD inhibitor, carbenoxolone (100 mg every 8 h for 7 d). d4F and its metabolites were measured in plasma and urine during a steady state infusion. Inhibition of 11{beta}HSDs by carbenoxolone was measured by increased steady state concentrations of d4F (41 {+/-} 5.1 vs. 48 {+/-} 7.7 nM; P < 0.05) and a fall in the rate of appearance of d3F (P < 0.05). 11{beta}HSD1 reductase activity could be measured specifically as conversion of d3E to d3F (28 {+/-} 4.2 vs. 17 {+/-} 3.1 nM; P < 0.05), whereas 11{beta}HSD2 could be measured by initial rates of appearance of d3E or from urinary ratios of d4F/(d3E + d3F) (0.73 {+/-} 0.06 vs. 1.02 {+/-} 0.03; P < 0.05). This technique offers a significant advance in the methods available to measure turnover in 11{beta}HSDs and isozymes of 11{beta}HSDs in vivo in human studies, and this study confirms that carbenoxolone inhibits both isozymes of 11{beta}HSD. 10.1210/jc.87.1.277 KW - 11bhsd1 KW - 11bhsd2 KW - cortisol KW - isotope KW - tracer AU - Andrew, Ruth AU - Smith, Kenneth AU - Jones, Gregory AU - Walker, Brian PY - 2002/01/01/ UR - http://dx.doi.org/10.1210/jc.87.1.277 ER -