Activation of MAPK in proximal tubule cells from spontaneously hypertensive and control Wistar-Kyoto rats

Abstract—The aim of this study was to test the hypothesis that differences exist in the activity and/or expression of mitogen-activated protein kinases (MAPKs) between spontaneously hypertensive rats (SHR) and control Wistar-Kyoto rats (WKY) and that these differences may account for the enhanced activity of the Na1/H1 exchanger (NHE) previously observed in the renal proximal tubule of SHR. Therefore, the activities of c-jun N-terminal kinase1 (JNK1), extracellular signal-regulated kinase1/2 (ERK1/2), and p38 were investigated. A reduced amount of ERK1 and JNK1 protein was found in renal cortex specimens of SHR as compared with WKY; however, their activities were the same. To study the cellular basis of this difference, immortalized proximal tubule cell lines were grown on Millicell-CM filter inserts where the cell lines organize as polarized monolayers with separate access to apical and basolateral compartments. Although basal JNK1 and ERK1/2 activities were not significantly different between WKY and SHR cells, anisomycin stimulated JNK1 activity in WKY cells more than in SHR cells (eg, at 15 minutes 300% versus 30%, respectively). Similarly, angiotensin II increased JNK1 and ERK1/2 activity in a time- and concentration-dependent manner in WKY cells but not in SHR cells. Western blot analyses showed a deficit in JNK1 and ERK1 protein in SHR (0.25 and 0.5, respectively, of the levels in WKY cells), although ERK2 and p38 protein levels were the same. These observations suggest that, although angiotensin II activates MAPKs and MAPKs have been shown to regulate NHE, this regulatory pathway is unlikely to account for the increased activity of NHE in the proximal tubular epithelium of SHR.