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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) Effect of chronic exposure to the selective 11β-HSD1 inhibitor                        PF-877423 on subcutaneous s–v cells. 11 β-HSD1 enzyme                        inhibition by 300 nM PF-877423 measured as the production of                        cortisol (day 6: 154±8 vs 5387±182; day 9:                        128±1 vs 5489±230; day 14: 174±18 vs                        4041±106; day 20: 409±27 vs 10443±78; day                        22: 330±7 vs 11218±193 pg/ml per                        24 h, mean±s.d.,                        P<0·001, n=3,                        E− or E+PF-877423-treated respectively). (B) Inhibition                        of lipid accumulation by 300 nM PF-877423 in subcutaneous                        stromal–vascular cells (day 16:                        0·25±0·03 vs                        0·20±0·01; day 20:                        0·3±0·02 vs                        0·20±0·01; day 22:                        0·27±0·01 vs                        0·19±0·01; OD (500/660 nm),                            n=3, E− or                        E+PF-877423-treated respectively), P values:                            **P<0·01,                        ***P<0·001.
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fig7: (A) Effect of chronic exposure to the selective 11β-HSD1 inhibitor PF-877423 on subcutaneous s–v cells. 11 β-HSD1 enzyme inhibition by 300 nM PF-877423 measured as the production of cortisol (day 6: 154±8 vs 5387±182; day 9: 128±1 vs 5489±230; day 14: 174±18 vs 4041±106; day 20: 409±27 vs 10443±78; day 22: 330±7 vs 11218±193 pg/ml per 24 h, mean±s.d., P<0·001, n=3, E− or E+PF-877423-treated respectively). (B) Inhibition of lipid accumulation by 300 nM PF-877423 in subcutaneous stromal–vascular cells (day 16: 0·25±0·03 vs 0·20±0·01; day 20: 0·3±0·02 vs 0·20±0·01; day 22: 0·27±0·01 vs 0·19±0·01; OD (500/660 nm), n=3, E− or E+PF-877423-treated respectively), P values: **P<0·01, ***P<0·001.

Mentions: Human subcutaneous s–v cells differentiated with E+PF-877423 had significantly lower 11β-HSD1 oxo-reductase activity compared with cells differentiated with E at any time point studied (day 6: 154±8 vs 5387±182; day 9: 128±1 vs 5489±230; day 14: 174±18 vs 4041±106; day 20: 409±27 vs 10443±78; day 22: 330±7 vs 11218±193 pg/ml per 24 h, mean±s.d., P<0·001, n=3, E− or E+PF-877423 treated respectively (Fig. 7A)). Lipid content in cells differentiated with E and PF-877423 was significantly lower than in cells differentiated with E only and similar to undifferentiated cells (day 16, 0·25±0·03 vs 0·20±0·01; day 20, 0·3±0·02 vs 0·20±0·01; day 22, 0·27±0·01 vs 0·19±0·01; OD (500/660 nm); mean±s.d., P<0·01, n=3, E− or E+PF-877423 treated respectively; Fig. 7B).


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) Effect of chronic exposure to the selective 11β-HSD1 inhibitor                        PF-877423 on subcutaneous s–v cells. 11 β-HSD1 enzyme                        inhibition by 300 nM PF-877423 measured as the production of                        cortisol (day 6: 154±8 vs 5387±182; day 9:                        128±1 vs 5489±230; day 14: 174±18 vs                        4041±106; day 20: 409±27 vs 10443±78; day                        22: 330±7 vs 11218±193 pg/ml per                        24 h, mean±s.d.,                        P<0·001, n=3,                        E− or E+PF-877423-treated respectively). (B) Inhibition                        of lipid accumulation by 300 nM PF-877423 in subcutaneous                        stromal–vascular cells (day 16:                        0·25±0·03 vs                        0·20±0·01; day 20:                        0·3±0·02 vs                        0·20±0·01; day 22:                        0·27±0·01 vs                        0·19±0·01; OD (500/660 nm),                            n=3, E− or                        E+PF-877423-treated respectively), P values:                            **P<0·01,                        ***P<0·001.
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fig7: (A) Effect of chronic exposure to the selective 11β-HSD1 inhibitor PF-877423 on subcutaneous s–v cells. 11 β-HSD1 enzyme inhibition by 300 nM PF-877423 measured as the production of cortisol (day 6: 154±8 vs 5387±182; day 9: 128±1 vs 5489±230; day 14: 174±18 vs 4041±106; day 20: 409±27 vs 10443±78; day 22: 330±7 vs 11218±193 pg/ml per 24 h, mean±s.d., P<0·001, n=3, E− or E+PF-877423-treated respectively). (B) Inhibition of lipid accumulation by 300 nM PF-877423 in subcutaneous stromal–vascular cells (day 16: 0·25±0·03 vs 0·20±0·01; day 20: 0·3±0·02 vs 0·20±0·01; day 22: 0·27±0·01 vs 0·19±0·01; OD (500/660 nm), n=3, E− or E+PF-877423-treated respectively), P values: **P<0·01, ***P<0·001.
Mentions: Human subcutaneous s–v cells differentiated with E+PF-877423 had significantly lower 11β-HSD1 oxo-reductase activity compared with cells differentiated with E at any time point studied (day 6: 154±8 vs 5387±182; day 9: 128±1 vs 5489±230; day 14: 174±18 vs 4041±106; day 20: 409±27 vs 10443±78; day 22: 330±7 vs 11218±193 pg/ml per 24 h, mean±s.d., P<0·001, n=3, E− or E+PF-877423 treated respectively (Fig. 7A)). Lipid content in cells differentiated with E and PF-877423 was significantly lower than in cells differentiated with E only and similar to undifferentiated cells (day 16, 0·25±0·03 vs 0·20±0·01; day 20, 0·3±0·02 vs 0·20±0·01; day 22, 0·27±0·01 vs 0·19±0·01; OD (500/660 nm); mean±s.d., P<0·01, n=3, E− or E+PF-877423 treated respectively; Fig. 7B).

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