Original ContributionAldosterone increases kidney tubule cell oxidants through calcium-mediated activation of NADPH oxidase and nitric oxide synthase
Section snippets
Materials
MDCK and LLC-PK1 cells were obtained from the American Type Culture Collection (Manassas, VA, USA). Cell culture media and reagents were obtained from PAA Laboratories GmbH (Pasching, Austria) and Invitrogen Life Technologies (Carlsbad, CA, USA), respectively. The primary antibody against p47phox (sc-14015) was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and α-tubulin (T6199) was purchased from Sigma (St. Louis, MO, USA). The oligonucleotide containing the consensus sequence
Aldosterone induces a calcium-mediated increase in ROS and RNS production via the mineralocorticoid receptor
Two kidney cell lines (LLC-PK1, MDCK) with different functional characteristics were chosen to study the mechanisms involved in the increased oxidant production associated with exposure to high aldosterone levels. LLC-PK1 cells are from pig kidney and resemble proximal tubule cells [29], and MDCK cells are from dog kidney and resemble distal tubule cells [30].
Mineralocorticoid and glucocorticoid receptors are expressed in LLC-PK1 [31] and MDCK cells (data not shown). To investigate if the
Discussion
We previously demonstrated that aldosterone is genotoxic in kidney tubule cells and causes the activation of NF-κB, with evidence that an increase in cellular oxidants in part triggers those effects [16], [31]. Both DNA damage and chronic NF-κB activation can lead to oncogenesis. Understanding the mechanism underlying aldosterone-induced oxidant production is essential in the development of therapies that could prevent the adverse effects of chronic aldosteronism, particularly the potential
Acknowledgments
This work was supported by grants from the University of California at Davis (USA) and by Deutsche Forschungsgemeinschaft Grant Schu 2367/1-1 (to N.S. and H.S.) (Germany). N.Q. was supported by a fellowship from the DAAD (Germany).
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