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Lang, F.; Klingel, K.; Wagner, C. A.; Stegen, C.; Warntges, S.; Friedrich, B.; Lanzendorfer, M.; Melzig, J.; Moschen, I.; Steuer, S.; Waldegger, S.; Sauter, M.; Paulmichl, M.; Gerke, V.; Risler, T.; Gamba, G.; Capasso, G.; Kandolf, R.; Hebert, S. C.; Massry, S. G.; Broer, S.
Proceedings of the National Academy of Sciences - PNAS, 07/2000, Letnik: 97, Številka: 14Journal Article
Transforming growth factor β (TGF-β ) has been shown to participate in the pathophysiology of diabetic complications. As shown most recently, TGF-β stimulates the expression of a distinct serine/threonine kinase (hSGK) which had previously been cloned as an early gene transcriptionally regulated by cell volume alterations. The present study was performed to elucidate transcription and function of hSGK in diabetic nephropathy. As shown by Northern blotting, an increase of extracellular glucose concentration increased hSGK mRNA levels in cultured cells, an effect qualitatively mimicked by osmotic cell shrinkage or treatment with TGF-β (2 μ g/liter), phorbol 12,13-didecanoate (1 μ M), or the Ca2+ionophore ionomycin (1 μ M) and blunted by high concentrations of nifedipine (10 and 100 μ M). In situ hybridization revealed that hSGK transcription was markedly enhanced in diabetic nephropathy, with particularly high expression in mesangial cells, interstitial cells, and cells in thick ascending limbs of Henle's loop and distal tubules. According to voltage clamp and tracer flux studies in Xenopus oocytes expressing the renal epithelial Na+channel ENaC or the mouse thick ascending limb Na+,K+,2Cl-cotransporter BSC-1, coexpression with hSGK stimulated ENaC and BSC-1 11-fold and 6-fold, respectively, effects reversed by kinase inhibitors staurosporine (1 μ M) and chelerythrine (1 μ M) and not elicited by inactive hSGK. In conclusion, excessive extracellular glucose concentrations enhance hSGK transcription, which in turn stimulates renal tubular Na+transport. These observations disclose an additional element in the pathophysiology of diabetic nephropathy.
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