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A novel selective 11beta-hydroxysteroid dehydrogenase type 1 inhibitor prevents human adipogenesis.

Bujalska IJ, Gathercole LL, Tomlinson JW, Darimont C, Ermolieff J, Fanjul AN, Rejto PA, Stewart PM - J. Endocrinol. (2008)

Bottom Line: Human cells were differentiated with 1.0 microM cortisol (F), or cortisone (E) with or without 100 nM of a highly selective 11beta-HSD1 inhibitor PF-877423. 11beta-HSD1 mRNA expression increased across adipocyte differentiation (P<0.001, n=4), which was paralleled by an increase in 11beta-HSD1 oxo-reductase activity (from nil on day 0 to 5.9+/-1.9 pmol/mg per h on day 16, P<0.01, n=7).In addition, cellular lipid content decreased significantly.These findings were confirmed in the primary cultures of human subcutaneous preadipocytes.

View Article: PubMed Central - PubMed

Affiliation: Division of Medical Sciences, The Medical School, Institute of Biomedical Research, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.

ABSTRACT
Glucocorticoid excess increases fat mass, preferentially within omental depots; yet circulating cortisol concentrations are normal in most patients with metabolic syndrome (MS). At a pre-receptor level, 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) activates cortisol from cortisone locally within adipose tissue, and inhibition of 11beta-HSD1 in liver and adipose tissue has been proposed as a novel therapy to treat MS by reducing hepatic glucose output and adiposity. Using a transformed human subcutaneous preadipocyte cell line (Chub-S7) and human primary preadipocytes, we have defined the role of glucocorticoids and 11beta-HSD1 in regulating adipose tissue differentiation. Human cells were differentiated with 1.0 microM cortisol (F), or cortisone (E) with or without 100 nM of a highly selective 11beta-HSD1 inhibitor PF-877423. 11beta-HSD1 mRNA expression increased across adipocyte differentiation (P<0.001, n=4), which was paralleled by an increase in 11beta-HSD1 oxo-reductase activity (from nil on day 0 to 5.9+/-1.9 pmol/mg per h on day 16, P<0.01, n=7). Cortisone enhanced adipocyte differentiation; fatty acid-binding protein 4 expression increased 312-fold (P<0.001) and glycerol-3-phosphate dehydrogenase 47-fold (P<0.001) versus controls. This was abolished by co-incubation with PF-877423. In addition, cellular lipid content decreased significantly. These findings were confirmed in the primary cultures of human subcutaneous preadipocytes. The increase in 11beta-HSD1 mRNA expression and activity is essential for the induction of human adipogenesis. Blocking adipogenesis with a novel and specific 11beta-HSD1 inhibitor may represent a novel approach to treat obesity in patients with MS.

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(A) PF-877423 inhibits 11β-HSD1 enzyme activity (dehydrogenase:                        12·4±1·0 vs                        0·2±0·01, % cortisol to cortisone                        conversion, and oxo-reductase: 34·7±0·6 vs                        0·4±0·1, % cortisone to cortisol                        conversion, mean±s.d.) as measured in HEK293T1 (HEK293                        cells stably transfected with human 11β-HSD type 1 cDNA),                        n=3 but not (B) 11β-HSD2 enzyme                        activity (63·6±4·0 vs                        62·2±4·4, % cortisol to cortisone                        conversion, mean±s.d., control versus                        PF-9=877423 respectively) as measured in HEK293T2 (cells stably                        transfected with human 11β-HSD type 2 cDNA),                        n=3. P values:                        **P<0·01,                        ***P<0·001.
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fig2: (A) PF-877423 inhibits 11β-HSD1 enzyme activity (dehydrogenase: 12·4±1·0 vs 0·2±0·01, % cortisol to cortisone conversion, and oxo-reductase: 34·7±0·6 vs 0·4±0·1, % cortisone to cortisol conversion, mean±s.d.) as measured in HEK293T1 (HEK293 cells stably transfected with human 11β-HSD type 1 cDNA), n=3 but not (B) 11β-HSD2 enzyme activity (63·6±4·0 vs 62·2±4·4, % cortisol to cortisone conversion, mean±s.d., control versus PF-9=877423 respectively) as measured in HEK293T2 (cells stably transfected with human 11β-HSD type 2 cDNA), n=3. P values: **P<0·01, ***P<0·001.

Mentions: 11β-HSD enzyme assays on HEK293T1 and HEK293T2 cells showed total abolition of dehydrogenase (12·4±1·0 vs 0·2±0·01, % cortisol to cortisone conversion, mean±s.d.) and oxo-reductase (34·7±0·6 vs 0·4±0·1, % cortisone to cortisol conversion, mean±s.d.) activities of 11β-HSD1 following incubation with 100 nM PF-877423 for 24 h (Fig. 2A), but PF-877423 had no effect on 11β-HSD2 activity (63·6±4·0 vs 62·2±4·4, % cortisol to cortisone conversion, mean±s.d., control versus PF-877423 respectively; Fig. 2B). No toxic effects of PF-877423 were observed up to 10 μM concentrations using a commercially available assay kit (CellTiter 96 Aqueous, Promega; data not shown).


A novel selective 11beta-hydroxysteroid dehydrogenase type 1 inhibitor prevents human adipogenesis.

Bujalska IJ, Gathercole LL, Tomlinson JW, Darimont C, Ermolieff J, Fanjul AN, Rejto PA, Stewart PM - J. Endocrinol. (2008)

(A) PF-877423 inhibits 11β-HSD1 enzyme activity (dehydrogenase:                        12·4±1·0 vs                        0·2±0·01, % cortisol to cortisone                        conversion, and oxo-reductase: 34·7±0·6 vs                        0·4±0·1, % cortisone to cortisol                        conversion, mean±s.d.) as measured in HEK293T1 (HEK293                        cells stably transfected with human 11β-HSD type 1 cDNA),                        n=3 but not (B) 11β-HSD2 enzyme                        activity (63·6±4·0 vs                        62·2±4·4, % cortisol to cortisone                        conversion, mean±s.d., control versus                        PF-9=877423 respectively) as measured in HEK293T2 (cells stably                        transfected with human 11β-HSD type 2 cDNA),                        n=3. P values:                        **P<0·01,                        ***P<0·001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC2315694&req=5

fig2: (A) PF-877423 inhibits 11β-HSD1 enzyme activity (dehydrogenase: 12·4±1·0 vs 0·2±0·01, % cortisol to cortisone conversion, and oxo-reductase: 34·7±0·6 vs 0·4±0·1, % cortisone to cortisol conversion, mean±s.d.) as measured in HEK293T1 (HEK293 cells stably transfected with human 11β-HSD type 1 cDNA), n=3 but not (B) 11β-HSD2 enzyme activity (63·6±4·0 vs 62·2±4·4, % cortisol to cortisone conversion, mean±s.d., control versus PF-9=877423 respectively) as measured in HEK293T2 (cells stably transfected with human 11β-HSD type 2 cDNA), n=3. P values: **P<0·01, ***P<0·001.
Mentions: 11β-HSD enzyme assays on HEK293T1 and HEK293T2 cells showed total abolition of dehydrogenase (12·4±1·0 vs 0·2±0·01, % cortisol to cortisone conversion, mean±s.d.) and oxo-reductase (34·7±0·6 vs 0·4±0·1, % cortisone to cortisol conversion, mean±s.d.) activities of 11β-HSD1 following incubation with 100 nM PF-877423 for 24 h (Fig. 2A), but PF-877423 had no effect on 11β-HSD2 activity (63·6±4·0 vs 62·2±4·4, % cortisol to cortisone conversion, mean±s.d., control versus PF-877423 respectively; Fig. 2B). No toxic effects of PF-877423 were observed up to 10 μM concentrations using a commercially available assay kit (CellTiter 96 Aqueous, Promega; data not shown).

Bottom Line: Human cells were differentiated with 1.0 microM cortisol (F), or cortisone (E) with or without 100 nM of a highly selective 11beta-HSD1 inhibitor PF-877423. 11beta-HSD1 mRNA expression increased across adipocyte differentiation (P<0.001, n=4), which was paralleled by an increase in 11beta-HSD1 oxo-reductase activity (from nil on day 0 to 5.9+/-1.9 pmol/mg per h on day 16, P<0.01, n=7).In addition, cellular lipid content decreased significantly.These findings were confirmed in the primary cultures of human subcutaneous preadipocytes.

View Article: PubMed Central - PubMed

Affiliation: Division of Medical Sciences, The Medical School, Institute of Biomedical Research, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.

ABSTRACT
Glucocorticoid excess increases fat mass, preferentially within omental depots; yet circulating cortisol concentrations are normal in most patients with metabolic syndrome (MS). At a pre-receptor level, 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) activates cortisol from cortisone locally within adipose tissue, and inhibition of 11beta-HSD1 in liver and adipose tissue has been proposed as a novel therapy to treat MS by reducing hepatic glucose output and adiposity. Using a transformed human subcutaneous preadipocyte cell line (Chub-S7) and human primary preadipocytes, we have defined the role of glucocorticoids and 11beta-HSD1 in regulating adipose tissue differentiation. Human cells were differentiated with 1.0 microM cortisol (F), or cortisone (E) with or without 100 nM of a highly selective 11beta-HSD1 inhibitor PF-877423. 11beta-HSD1 mRNA expression increased across adipocyte differentiation (P<0.001, n=4), which was paralleled by an increase in 11beta-HSD1 oxo-reductase activity (from nil on day 0 to 5.9+/-1.9 pmol/mg per h on day 16, P<0.01, n=7). Cortisone enhanced adipocyte differentiation; fatty acid-binding protein 4 expression increased 312-fold (P<0.001) and glycerol-3-phosphate dehydrogenase 47-fold (P<0.001) versus controls. This was abolished by co-incubation with PF-877423. In addition, cellular lipid content decreased significantly. These findings were confirmed in the primary cultures of human subcutaneous preadipocytes. The increase in 11beta-HSD1 mRNA expression and activity is essential for the induction of human adipogenesis. Blocking adipogenesis with a novel and specific 11beta-HSD1 inhibitor may represent a novel approach to treat obesity in patients with MS.

Show MeSH
Related in: MedlinePlus