Hypertonic stress activates glycogen synthase kinase 3β-mediated apoptosis of renal medullary interstitial cells, suppressing an NFκB-driven cyclooxygenase-2-dependent survival pathway

The survival of renal medullary interstitial cells (RMICs) requires their adaptation to rapid shifts in ambient tonicity normally occurring in the renal medulla. Previous studies determined that cyclooxygenase-2 (COX 2) activation is critical for this adaptation. The present studies find that these adaptive mechanisms are dampened by the simultaneous activation of an apoptotic pathway linked to a glycogen synthase kinase 3beta (GSK 3beta). Inhibition of GSK 3 by LiCl or specific small molecule GSK inhibitors increased RMIC survival following hypertonic stress, and transduction of RMICs with a constitutively active GSK 3beta (AdGSK 3betaA9) significantly increased apoptosis, consistent with a proapoptotic role of GSK 3beta. Following GSK 3beta inhibition, increased survival was accompanied by increased COX 2 expression and COX 2 reporter activity. In contrast, GSK 3beta overexpression reduced COX 2 reporter activity. Importantly, enhanced RMIC survival produced by GSK 3beta inhibition was completely dependent on COX 2 because it was abolished by a COX 2-specific inhibitor, SC58236. The signaling pathway by which GSK 3beta suppresses COX 2 expression was then explored. GSK 3beta inhibition increased both NFkappaB and beta-catenin activity associated with decreased IkappaB and increased beta-catenin levels. The increase in COX 2 following GSK 3beta inhibition was entirely blocked by NFkappaB inhibition using mutant IkappaB adenovirus. However, adenoviral overexpression of beta-catenin did not increase COX 2 levels. These findings suggest that GSK 3beta negatively regulates COX 2 expression and that GSK 3beta inhibitors protect RMICs from hypertonic stress via induction of NFkappaB-COX 2-dependent pathway.