The interdependence of EGF-R and SGK-1 in fibronectin expression in primary kidney cortical fibroblast cells

Background: Epidermal growth factor (EGF) has been shown to play a role in the nephromegaly and enhanced sodium reabsorption observed in diabetic nephropathy. This is recognized to be dependent on activation of serine threonine glucocorticoid kinase-1 (SGK-1). However, the roles of EGF and SGK-1 in renal fibrogenesis observed under high glucose conditions have not been established. Methods: Primary cultures of human cortical fibroblasts (CFs) were used as the model in which to study the dependent and independent effects of high glucose, EGF and SGK-1 on the expression of the extracellular matrix protein (ECM) fibronectin. Wild type CFs expressing SGK-1, or cells in which SGK-1 was effectively silenced using siRNA methodology, were exposed to normal (5 mM) or high (25 mM) glucose, or EGF (10 ng/ml) for 48 hr and fibronectin assessed. The role of the EGF-receptor and its relationship to SGK-1 signaling was studied using concurrent treatment with PKI166, a specific inhibitor of EGF-receptor. Results: Exposure of CF to high glucose and EGF increased phosphorylated EGF-R, SGK-1, and fibronectin expression in wildtype cells. Inhibition of the EGF-R reduced SGK-1 and fibronectin expression in control, and following exposure to EGF and high glucose conditions. In cells in which SGK-1 was silenced, fibronectin was reduced and there was no significant increase in pEGF-R, suggesting that SGK-1 is downstream of the EGF-R and negatively inhibits EGF-R activation. Conclusion: These results suggest that high glucose induced fibronectin expression is mediated through the EGF-R and downstream expression of SGK-1. (C) 2007 Elsevier Ltd. All rights reserved.