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J Nephrol. 2013 May-Jun;26(3):510-6. doi: 10.5301/jn.5000175. Epub 2012 Aug 7.

Albumin upregulates eNOS mRNA through ETRA/B in human proximal tubular epithelial cells.

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1
Department of Biology, University of Patras, Patras, Greece. katsopan@upatras.gr

Abstract

BACKGROUND:

Proximal tubular cells respond to proteinuria by expressing several cytokines and inflammatory molecules that induce interstitial fibrosis. Increased attention has been drawn toward the systems of endothelin (ET) and nitric oxide (NO). This work contributes to the elucidation of the interplay between these two systems in proximal tubular epithelial cells (PTECs) after exposure in proteinuric conditions.

METHODS:

HK-2 cells, a human PTEC line, were incubated with albumin, simulating proteinuric conditions. Cells were then lysed and either total RNA was isolated or whole cell extracts were prepared. PreproET-1, ET receptors (ETRA and ETRB) and NO synthases (eNOS, iNOS) mRNA accumulation was estimated by RT-PCR, and proteins by Western blot analysis. NO production was assessed using Griess reaction. Furthermore, we treated HK-2 cells with NO donor sodium nitroprusside, NO inhibitor L-NAME, ETRA inhibitor BQ123, ETRB inhibitor BQ788 and purified ET-1, and investigated the potential interplay between albumin-induced stimulation of NO or ET-1 systems.

RESULTS:

We found that albumin upregulates preproET-1, ETRA, ETRB, eNOS and iNOS mRNA as well as protein and stimulates NO production. Additionally, we recorded an ETRA/B dependent regulation of albumin-induced eNOS expression.

CONCLUSIONS:

For the first time an in vitro albumin-induced ET-1 and NO interplay was revealed.

PMID:
22878978
DOI:
10.5301/jn.5000175
[Indexed for MEDLINE]
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