Effects of cyclosporine A on Na,K-ATPase expression in the renal epithelial cell line NBL-1

Abstract

The bovine renal epithelial cell line NBL-1 has been used to monitor the effects of cyclosporine A (CsA) on Na+,K(+)-ATPase activity and expression. CsA at two single doses (0.6 mg/liter and 2.5 mg/liter) inhibits the ouabain-sensitive component of Rb+ uptake, assumed to be Na+,K(+)-ATPase, but increases the low activity of a furosemide-sensitive component corresponding to a Na+/K+/Cl- cotransporter. CsA addition also induces a slight decrease of alpha 1 subunit mRNA levels, without altering the already low beta 1 subunit mRNA amounts. Hypertonic treatment of NBL-1 cells leads to a significant increase in both Na+,K(+)-ATPase activity and alpha 1 subunit mRNA amounts, but does not modify beta 1 subunit mRNA levels. The differential response of the alpha 1 and beta 1 subunit genes may explain why hypertonic treatment does not result in higher alpha 1 protein expression, and supports the view that increased activity relies upon post-translational events, despite the likely transcriptional activation of the alpha 1 subunit gene. The addition of CsA does not alter the hypertonicity-mediated increase of Na+,K(+)-ATPase activity but blocks the accumulation of alpha 1 subunit mRNA. In conclusion, CsA may compromise the ion handling by renal cells as a result of the inhibition of basal Na+,K(+)-ATPase activity and the stimulation of Na+/K+/Cl- cotransport activity. Moreover, this is the first report showing that CsA may affect the long-term adaptation of the pump by altering its subunit gene expression.