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Tumor necrosis factor alpha and interleukin 1beta enhance the cortisone/cortisol shuttle.

Escher G, Galli I, Vishwanath BS, Frey BM, Frey FJ - J. Exp. Med. (1997)

Bottom Line: Since we found that glomerular mesangial cells (GMC) express 11beta-hydroxysteroid dehydrogenase 1 (11beta-OHSD1), which interconverts 11-keto glucocorticosteroids into 11beta-hydroxy glucocorticosteroids (cortisone/cortisol shuttle), we explored whether 11beta-OHSD1 determines the antiinflammatory effect of glucocorticosteroids.Thus, we conclude that 11beta-OHSD1 controls access of 11beta-hydroxy glucocorticosteroids and 11-keto glucocorticosteroids to glucocorticoid receptors and thus determines the anti-inflammatory effect of glucocorticosteroids.IL-1beta and TNF-alpha upregulate specifically the reductase activity of 11beta-OHSD1 and counterbalance by that mechanism their own proinflammatory effect.

View Article: PubMed Central - PubMed

Affiliation: Division of Nephrology, University Hospital of Berne, 3010 Berne, Switzerland.

ABSTRACT
Endogenously released or exogenously administered glucocorticosteroids are relevant hormones for controlling inflammation. Only 11beta-hydroxy glucocorticosteroids, but not 11-keto glucocorticosteroids, activate glucocorticoid receptors. Since we found that glomerular mesangial cells (GMC) express 11beta-hydroxysteroid dehydrogenase 1 (11beta-OHSD1), which interconverts 11-keto glucocorticosteroids into 11beta-hydroxy glucocorticosteroids (cortisone/cortisol shuttle), we explored whether 11beta-OHSD1 determines the antiinflammatory effect of glucocorticosteroids. GMC exposed to interleukin (IL)-1beta or tumor necrosis factor alpha (TNF-alpha) release group II phospholipase A2 (PLA2), a key enzyme producing inflammatory mediators. 11beta-hydroxy glucocorticosteroids inhibited cytokine-induced transcription and release of PLA2 through a glucocorticoid receptor-dependent mechanism. This inhibition was enhanced by inhibiting 11beta-OHSD1. Interestingly, 11-keto glucocorticosteroids decreased cytokine-induced PLA2 release as well, a finding abrogated by inhibiting 11beta-OHSD1. Stimulating GMC with IL-1beta or TNF-alpha increased expression and reductase activity of 11beta-OHSD1. Similarly, this IL-1beta- and TNF-alpha-induced formation of active 11beta-hydroxy glucocorticosteroids from inert 11-keto glucocorticosteroids by the 11beta-OHSD1 was shown in the Kiki cell line that expresses the stably transfected bacterial beta-galactosidase gene under the control of a glucocorticosteroids response element. Thus, we conclude that 11beta-OHSD1 controls access of 11beta-hydroxy glucocorticosteroids and 11-keto glucocorticosteroids to glucocorticoid receptors and thus determines the anti-inflammatory effect of glucocorticosteroids. IL-1beta and TNF-alpha upregulate specifically the reductase activity of 11beta-OHSD1 and counterbalance by that mechanism their own proinflammatory effect.

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(Top) IL-1β and  TNF-α increase steady-state  11β-OHSD1 mRNA in GMC.  The mean (± SEM, n = 6) incorporation of [α32P]-dCTP  (cpm) into cDNA of 11β-OHSD1 after incubation of  GMC with TNF-α and/or IL-1β  was standardized to that of  GAPDH mRNA. (Bottom) IL-1β  and TNF-α increase 11β-OHSD1  protein levels in GMC. Western  blot analysis of 11β-OHSD1 in  GMC stimulated with and without TNF-α and/or IL-1β. A,  11β-OHSD1 from COS cells  transfected with the cDNA of  11β-OHSD1; B, GMC; C,  GMC with 5 nM of TNF-α;  D, GMC with 5 nM of IL-1β; E,  GMC with 5 nM of TNF-α and  5 nM of IL-1β. Quantification  of 11β-OHSD1 protein by densitometry analysis of the Western blot.  Without cytokines, no 11β-OHSD1 protein was detected. Both IL-1β  and TNF-α increased the relative transmission from a signal of below the  detection limit to a value of 2–4.
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Figure 7: (Top) IL-1β and TNF-α increase steady-state 11β-OHSD1 mRNA in GMC. The mean (± SEM, n = 6) incorporation of [α32P]-dCTP (cpm) into cDNA of 11β-OHSD1 after incubation of GMC with TNF-α and/or IL-1β was standardized to that of GAPDH mRNA. (Bottom) IL-1β and TNF-α increase 11β-OHSD1 protein levels in GMC. Western blot analysis of 11β-OHSD1 in GMC stimulated with and without TNF-α and/or IL-1β. A, 11β-OHSD1 from COS cells transfected with the cDNA of 11β-OHSD1; B, GMC; C, GMC with 5 nM of TNF-α; D, GMC with 5 nM of IL-1β; E, GMC with 5 nM of TNF-α and 5 nM of IL-1β. Quantification of 11β-OHSD1 protein by densitometry analysis of the Western blot. Without cytokines, no 11β-OHSD1 protein was detected. Both IL-1β and TNF-α increased the relative transmission from a signal of below the detection limit to a value of 2–4.

Mentions: Since it was shown previously that IL-1β increased corticosteroid levels, the hypothesis was tested whether IL-1β and TNF-α regulate the 11β-OHSD1 (33, 34). For that purpose, GMC were incubated with TNF-α, IL-1β, or a combination of both, and the mRNA content was quantified by RT-PCR for 11β-OHSD1 (Fig. 7, top). Both IL-1β and TNF-α only slightly enhanced the mRNA content in GMC. The combination of both, in contrast, yielded a strong additive effect (Fig. 7, top). The increased mRNA content of GMC was reflected by a sharp increase in the amount of 11β-OHSD1 protein (Fig. 7, bottom). The cytokine-induced increase of protein levels was more marked than the effect on steady-state mRNA levels, suggesting that the modulation may take place mainly at the translational level. When the specific activity of 11β-OHSD1 was analyzed, no change was seen with respect to oxidation (results not shown). However, the reductive activity of 11β-OHSD1 increased as a function of the concentrations of IL-1β and/or TNF-α (Fig. 8). The combination of TNF-α and IL-1β stimulated the reductive activity more than either of the cytokines added alone; this was shown when either increasing concentrations of TNF-α or IL-1β were used, or when a fixed amount of TNF-α with an increasing amount of IL-1β were given to the cell cultures (Fig. 8). A similar pattern was observed when the specific activity (percentage of conversion of dehydrocorticosterone to corticosterone per milligram protein per 4-h period) was calculated (data not shown). To exclude that the increased reductive activity after stimulation with cytokines was due to an increased availability of cofactors, incubations of stimulated GMC with dehydrocorticosterone in the presence of 5 mM NADPH were performed. The results from these studies revealed that the increased reductase activity induced by cytokines was not attributable to a higher availability of NADPH (data not shown).


Tumor necrosis factor alpha and interleukin 1beta enhance the cortisone/cortisol shuttle.

Escher G, Galli I, Vishwanath BS, Frey BM, Frey FJ - J. Exp. Med. (1997)

(Top) IL-1β and  TNF-α increase steady-state  11β-OHSD1 mRNA in GMC.  The mean (± SEM, n = 6) incorporation of [α32P]-dCTP  (cpm) into cDNA of 11β-OHSD1 after incubation of  GMC with TNF-α and/or IL-1β  was standardized to that of  GAPDH mRNA. (Bottom) IL-1β  and TNF-α increase 11β-OHSD1  protein levels in GMC. Western  blot analysis of 11β-OHSD1 in  GMC stimulated with and without TNF-α and/or IL-1β. A,  11β-OHSD1 from COS cells  transfected with the cDNA of  11β-OHSD1; B, GMC; C,  GMC with 5 nM of TNF-α;  D, GMC with 5 nM of IL-1β; E,  GMC with 5 nM of TNF-α and  5 nM of IL-1β. Quantification  of 11β-OHSD1 protein by densitometry analysis of the Western blot.  Without cytokines, no 11β-OHSD1 protein was detected. Both IL-1β  and TNF-α increased the relative transmission from a signal of below the  detection limit to a value of 2–4.
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Figure 7: (Top) IL-1β and TNF-α increase steady-state 11β-OHSD1 mRNA in GMC. The mean (± SEM, n = 6) incorporation of [α32P]-dCTP (cpm) into cDNA of 11β-OHSD1 after incubation of GMC with TNF-α and/or IL-1β was standardized to that of GAPDH mRNA. (Bottom) IL-1β and TNF-α increase 11β-OHSD1 protein levels in GMC. Western blot analysis of 11β-OHSD1 in GMC stimulated with and without TNF-α and/or IL-1β. A, 11β-OHSD1 from COS cells transfected with the cDNA of 11β-OHSD1; B, GMC; C, GMC with 5 nM of TNF-α; D, GMC with 5 nM of IL-1β; E, GMC with 5 nM of TNF-α and 5 nM of IL-1β. Quantification of 11β-OHSD1 protein by densitometry analysis of the Western blot. Without cytokines, no 11β-OHSD1 protein was detected. Both IL-1β and TNF-α increased the relative transmission from a signal of below the detection limit to a value of 2–4.
Mentions: Since it was shown previously that IL-1β increased corticosteroid levels, the hypothesis was tested whether IL-1β and TNF-α regulate the 11β-OHSD1 (33, 34). For that purpose, GMC were incubated with TNF-α, IL-1β, or a combination of both, and the mRNA content was quantified by RT-PCR for 11β-OHSD1 (Fig. 7, top). Both IL-1β and TNF-α only slightly enhanced the mRNA content in GMC. The combination of both, in contrast, yielded a strong additive effect (Fig. 7, top). The increased mRNA content of GMC was reflected by a sharp increase in the amount of 11β-OHSD1 protein (Fig. 7, bottom). The cytokine-induced increase of protein levels was more marked than the effect on steady-state mRNA levels, suggesting that the modulation may take place mainly at the translational level. When the specific activity of 11β-OHSD1 was analyzed, no change was seen with respect to oxidation (results not shown). However, the reductive activity of 11β-OHSD1 increased as a function of the concentrations of IL-1β and/or TNF-α (Fig. 8). The combination of TNF-α and IL-1β stimulated the reductive activity more than either of the cytokines added alone; this was shown when either increasing concentrations of TNF-α or IL-1β were used, or when a fixed amount of TNF-α with an increasing amount of IL-1β were given to the cell cultures (Fig. 8). A similar pattern was observed when the specific activity (percentage of conversion of dehydrocorticosterone to corticosterone per milligram protein per 4-h period) was calculated (data not shown). To exclude that the increased reductive activity after stimulation with cytokines was due to an increased availability of cofactors, incubations of stimulated GMC with dehydrocorticosterone in the presence of 5 mM NADPH were performed. The results from these studies revealed that the increased reductase activity induced by cytokines was not attributable to a higher availability of NADPH (data not shown).

Bottom Line: Since we found that glomerular mesangial cells (GMC) express 11beta-hydroxysteroid dehydrogenase 1 (11beta-OHSD1), which interconverts 11-keto glucocorticosteroids into 11beta-hydroxy glucocorticosteroids (cortisone/cortisol shuttle), we explored whether 11beta-OHSD1 determines the antiinflammatory effect of glucocorticosteroids.Thus, we conclude that 11beta-OHSD1 controls access of 11beta-hydroxy glucocorticosteroids and 11-keto glucocorticosteroids to glucocorticoid receptors and thus determines the anti-inflammatory effect of glucocorticosteroids.IL-1beta and TNF-alpha upregulate specifically the reductase activity of 11beta-OHSD1 and counterbalance by that mechanism their own proinflammatory effect.

View Article: PubMed Central - PubMed

Affiliation: Division of Nephrology, University Hospital of Berne, 3010 Berne, Switzerland.

ABSTRACT
Endogenously released or exogenously administered glucocorticosteroids are relevant hormones for controlling inflammation. Only 11beta-hydroxy glucocorticosteroids, but not 11-keto glucocorticosteroids, activate glucocorticoid receptors. Since we found that glomerular mesangial cells (GMC) express 11beta-hydroxysteroid dehydrogenase 1 (11beta-OHSD1), which interconverts 11-keto glucocorticosteroids into 11beta-hydroxy glucocorticosteroids (cortisone/cortisol shuttle), we explored whether 11beta-OHSD1 determines the antiinflammatory effect of glucocorticosteroids. GMC exposed to interleukin (IL)-1beta or tumor necrosis factor alpha (TNF-alpha) release group II phospholipase A2 (PLA2), a key enzyme producing inflammatory mediators. 11beta-hydroxy glucocorticosteroids inhibited cytokine-induced transcription and release of PLA2 through a glucocorticoid receptor-dependent mechanism. This inhibition was enhanced by inhibiting 11beta-OHSD1. Interestingly, 11-keto glucocorticosteroids decreased cytokine-induced PLA2 release as well, a finding abrogated by inhibiting 11beta-OHSD1. Stimulating GMC with IL-1beta or TNF-alpha increased expression and reductase activity of 11beta-OHSD1. Similarly, this IL-1beta- and TNF-alpha-induced formation of active 11beta-hydroxy glucocorticosteroids from inert 11-keto glucocorticosteroids by the 11beta-OHSD1 was shown in the Kiki cell line that expresses the stably transfected bacterial beta-galactosidase gene under the control of a glucocorticosteroids response element. Thus, we conclude that 11beta-OHSD1 controls access of 11beta-hydroxy glucocorticosteroids and 11-keto glucocorticosteroids to glucocorticoid receptors and thus determines the anti-inflammatory effect of glucocorticosteroids. IL-1beta and TNF-alpha upregulate specifically the reductase activity of 11beta-OHSD1 and counterbalance by that mechanism their own proinflammatory effect.

Show MeSH
Related in: MedlinePlus