Abstract

Our previous study demonstrated that the GR is expressed in the human kidney glomerulus. The function of the GR of glomerular cells might be affected by the concentration of intracellular glucocorticoids, which is modulated by 11beta-hydroxysteroid dehydrogenase type 2 (11betaHSD2). Because the expression of 11betaHSD2 in the glomerular cells remains unclear, we used competitive RT-PCR and immunoblotting to detect the expression of 11betaHSD2 mRNA and protein in isolated human glomeruli, in whole kidney cortex as a positive control, and in a human glomerular visceral epithelial cell line. 11betaHSD2 mRNA was detected in all samples. Specific antihuman 11betaHSD2 antibody recognized a single band at 41 kDa, consistent with the molecular mass of human 11betaHSD2, in the samples of the isolated glomeruli and whole kidney cortex. Furthermore, definite 11betaHSD2 enzymatic activity was also determined with the sample of isolated glomeruli. We also performed immunohistochemistry by light and electron microscopy to determine the cellular and subcellular localization of 11betaHSD2 in the human glomeruli. Immunoreactivity of the enzyme was clearly observed in the glomerular visceral epithelial cells and endothelial cells as well as in the distal convoluted tubules and collecting ducts. The subcellular localization of 11betaHSD2 was shown to be endoplasmic reticulum. These results suggest that 11betaHSD2 might play a crucial role in modulating the intracellular concentration of glucocorticoids in human glomerular cells.

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