RhoA/ROCK inhibition attenuates endothelin-1-induced glomerulopathy in the rats

Abstract

Endothelin-1 (ET-1) contributes to the development of kidney diseases. However, the underlying molecular mechanism is largely undefined. Here we sought to investigate the potential role of ET-1 receptors, ET_A and ET_B in the regulation of increased glomerular permeability and underlying signaling pathways post-ET-1 infusion. Male Sprague-Dawley rats were infused with ET-1 (2 pmol/kg per minute, i.v.) for four weeks, and the effect on glomerular permeability to albumin (P_alb) and albuminuria was measured. The selective ROCK-1/2 inhibitor, Y-27632, was administered to a separate group of rats to determine its effect on ET-1-induced P_alb and albuminuria. The role of ET_A and ET_B receptors in regulating RhoA/ROCK activity was determined by incubating isolated glomeruli from normal rats with ET-1 and with selective ET_A and ET_B receptor antagonists. ET-1 infusion for four weeks significantly elevated P_alb and albuminuria. Y-27632 significantly reduced the elevation of P_alb and albuminuria. The activities of both RhoA and ROCK-1/2 were increased by ET-1 infusion. Selective ET_B receptor antagonism had no effect on the elevated activity of both RhoA and ROCK-1/2 enzymes. Selective ET_A receptor and combined ET_A/ET_B receptors blockade restored the activity of RhoA and ROCK-1/2 to normal levels. In addition, chronic ET-1 infusion increased the levels of glomerular inflammatory and fibrotic markers. These effects were all attenuated in rats following ROCK-1/2 inhibition. These observations suggest that ET-1 contributes to increased albuminuria, inflammation, and fibrosis by modulating the activity of the ET_A-RhoA/ROCK-1/2 pathway. Selective ET_A receptor blockade may represent a potential therapeutic strategy to limit glomerular injury and albuminuria in kidney disease.

Keywords: Albuminuria; Chronic kidney disease (CKD); Endothelin receptor antagonists (ERAs); Endothelin-1; Glomerular fibrosis; Glomerular inflammation; Glomerular permeability to albumin (P(alb)); ROCK-1/2; RhoA.