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Articles in PresS, published online ahead of print December 10, 2002
Am J Physiol Renal Physiol, 10.1152/ajprenal.00221.2002
Submitted on June 10, 2002
Accepted on December 3, 2002
1 Department of Nephrology, Tel Aviv Sourasky Medical Center, Tel Aviv, Israel
* To whom correspondence should be addressed. E-mail: nmdri{at}netvision.net.il.
The decrease in GFR which is characteristic of sepsis, has been shown to result from inhibition of the glomerular endothelial nitric oxide synthase (NOS) by nitric oxide (NO) generated from the inducible isoform of NOS (iNOS). Although, L-arginine is the sole precursor for NO biosynthesis, it's intracellular availability in glomeruli from septic animals has never been investigated. Arginine uptake was measured in freshly harvested glomeruli from the following experimental groups: 1. Untreated rats; 2. Rats pre- treated with lipopolysaccharide (LPS) (4 mg/kg BW, 4 hours prior to experiments), 3. Rats treated with LPS as above, with either L-NIL [L-N6-(1-Iminoethyl)lysine hydrochloride], a selective iNOS antagonist, or 7NI (7-nitriindazole a selective bNOS antagonist) 4. L-NIL only. Both glomeular and mesangial arginine transport characteristics were found compatible with a y+ system. Arginine uptake was augmented in glomeruli from LPS treated rats. Treatment with L-NIL completely abolished this effect while L-NIL alone had no effect. Similar results were obtained when primary cultures of rat mesangial cells were pre-incubated with LPS (10 µmg/ml for 24 hours) with or without L-NIL. Using RT-PCR, we found that in vivo administration of LPS resulted in a significant increase in glomerular cationic amino acid 2 (CAT-2) mRNA expression while CAT-1 mRNA was undetected. Northern blotting further confirmed a significant increase in glomerular CAT-2 by LPS. In mesangial cells, both the expression of CAT-1 and CAT-2 mRNA were augmented following incubation with LPS. Conclusions: In vivo administration of LPS augments glomerular arginine transport through upregulation of steady state CAT-2 mRNA while down regulating CAT-1 mRNA. These results may correspond to the changes in glomerular iNOS and eNOS activity in sepsis.
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