BACKGROUND: Glucocorticoids are efficiently used as antiinflammatory and immunosuppressive therapies of renal diseases. However, long-term treatment often is associated with net changes in the turnover of extracellular matrix (ECM) components. METHODS: We examined the impact of glucocorticoids on cAMP-triggered expression of tissue plasminogen activator (tPA), a protease prominently involved in glomerular ECM turnover. RESULTS: By ELISA, the db-cAMP-mediated increase in extracellular tPA activity secreted by mesangial cells (MC) was markedly reduced in the presence of 100 nmol/L dexamethasone. The decrease of enzymatic activity was accompanied by an attenuation of tPA expression, as shown by Northern blot analysis. Furthermore, dexamethasone increased the steady-state mRNA level of the tPA-inhibitor 1 (PAI-1), thereby providing an additional mode of regulation of tPA activity. Mutational analysis revealed that the inhibition of tPA expression was localized within the proximal 2.3 kb of the 5'-flanking region of the rat tPA gene and critically depended on a cAMP response element (CRE) at position -185. EMSA demonstrated that binding to this CRE was affected by dexamethasone, since the db-cAMP-caused DNA binding of CREB and C/EBPbeta-immunopositive complexes was substantially reduced by dexamethasone. In parallel, dexamethasone decreased the nuclear abundance of db-cAMP-induced C/EBPbeta and phosphorylated CREB protein without affecting the total level of either transcription factor. CONCLUSIONS: Suppression of cAMP-stimulated tPA expression by glucocorticoids occurs by interference with CREB and C/EBPbeta, the major transcription factors mediating cAMP responses. These observations may provide the molecular basis for the sclerotic processes within the glomerulus often complicating chronic glucocorticoid treatment.
BACKGROUND: Glucocorticoids are efficiently used as antiinflammatory and immunosuppressive therapies of renal diseases. However, long-term treatment often is associated with net changes in the turnover of extracellular matrix (ECM) components. METHODS: We examined the impact of glucocorticoids on cAMP-triggered expression of tissue plasminogen activator (tPA), a protease prominently involved in glomerular ECM turnover. RESULTS: By ELISA, the db-cAMP-mediated increase in extracellular tPA activity secreted by mesangial cells (MC) was markedly reduced in the presence of 100 nmol/L dexamethasone. The decrease of enzymatic activity was accompanied by an attenuation of tPA expression, as shown by Northern blot analysis. Furthermore, dexamethasone increased the steady-state mRNA level of the tPA-inhibitor 1 (PAI-1), thereby providing an additional mode of regulation of tPA activity. Mutational analysis revealed that the inhibition of tPA expression was localized within the proximal 2.3 kb of the 5'-flanking region of the rattPA gene and critically depended on a cAMP response element (CRE) at position -185. EMSA demonstrated that binding to this CRE was affected by dexamethasone, since the db-cAMP-caused DNA binding of CREB and C/EBPbeta-immunopositive complexes was substantially reduced by dexamethasone. In parallel, dexamethasone decreased the nuclear abundance of db-cAMP-induced C/EBPbeta and phosphorylated CREB protein without affecting the total level of either transcription factor. CONCLUSIONS: Suppression of cAMP-stimulated tPA expression by glucocorticoids occurs by interference with CREB and C/EBPbeta, the major transcription factors mediating cAMP responses. These observations may provide the molecular basis for the sclerotic processes within the glomerulus often complicating chronic glucocorticoid treatment.
Authors: Lili Li; Katherine Howell; Michelle Sands; Mark Banahan; Stephen Frohlich; Simon C Rowan; Roisín Neary; Donal Ryan; Paul McLoughlin Journal: PLoS One Date: 2013-12-09 Impact factor: 3.240