Somatostatin increases glucocorticoid binding and signaling in macrophages by blocking the calpain-specific cleavage of Hsp 90.
Somatostatin has direct anti-inflammatory actions and participates in the anti-inflammatory actions of glucocorticoids, but the mechanisms underlying this regulation remain poorly understood. The objective of this study was to evaluate whether somatostatin increases glucocorticoid responsiveness by up-regulating glucocorticoid receptor (GR) expression and signaling. Somatostatin promoted a time- and dose-dependent increase in [(3)H]dexamethasone binding to RAW 264.7 macrophages. Cell exposure to 10 nM somatostatin for 18 h promoted a 2-fold increase in the number of GR sites per cell without significant modification of the affinity. Analysis of GR heterocomplex components demonstrated that somatostatin increased the level of heat shock protein (Hsp) 90, whereas the level of GR remained almost unchanged. The increase in Hsp 90 was associated with a decrease in the cleavage of its carboxyl-terminal domain. Evidence for the involvement of calpain inhibition in this process was obtained by the demonstration that 1) somatostatin induced a dose-dependent decrease in calpain activity and 2) calpain inhibitors, calpain inhibitor I and calpeptin, both abolished the cleavage of Hsp 90 and induced a dose-dependent increase in [(3)H]dexamethasone binding. Increases in glucocorticoid binding after somatostatin treatment were associated with similar increases in the ability of GR to transactivate a minimal promoter containing two glucocorticoid response elements (GRE) and to interfere with the activation of nuclear factor-kappaB (NF-kappaB). Thus, the present findings indicate that somatostatin increases glucocorticoid binding and signaling by limiting the calpain-specific cleavage of GR-associated Hsp 90. This mechanism may represent a novel target for intervention to increase glucocorticoid responsiveness.
Bellocq
A
A
Unité INSERM 489, Hôpital Tenon, Paris, France.
Doublier
S
S
Suberville
S
S
Perez
J
J
Escoubet
B
B
Fouqueray
B
B
Puyol
D R
DR
Baud
L
L
D000818:Animals; D002154:Calpain / Q000037:antagonists & inhibitors*; D002460:Cell Line; D004247:DNA / Q000378:metabolism; D003907:Dexamethasone / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D013004:Somatostatin / Q000494:pharmacology*
D018841:HSP90 Heat-Shock Proteins; D011965:Receptors, Glucocorticoid; D013004:Somatostatin; D003907:Dexamethasone; D004247:DNA; D002154:Calpain