Inducible nitric oxide synthase inhibitors reduce urinary markers of systemic oxidant stress in murine proliferative lupus nephritis.
BACKGROUND
Proliferative lupus nephritis (PLN) is characterized by increased expression of inducible nitric oxide (NO) synthase (iNOS). Inhibition of iNOS with NG-monomethyl L-arginine (L-NMMA) abrogates renal disease in two models of murine PLN, but the mechanism of this effect is unknown. Reactive oxygen species have both direct and indirect pathogenic effects in inflammatory lesions and are therefore potentially an important therapeutic target in PLN. We hypothesized that inhibition of iNOS activity would reduce ROS production in murine PLN.


METHODS
A dose escalation of L-NMMA (0, 20, 100, and 500 mg/kg/day) was performed in New Zealand Black x New Zealand White F1 (NZB/W) mice with active renal disease. Twenty-four-hour urine nitrate + nitrite (NOX) was measured with a chemiluminescence NO analyzer. Twenty-four-hour urine 8-isoprostane F2alpha (8-iso-PGF2alpha) was measured by gas chromatography-negative ion chemical ionization mass spectrometry. MRL-MpJFASlpr (MRL/lpr) and NZB/W mice were divided into three groups and given either L-NMMA, L-N6-iminoethyl-lysine (L-NIL), or distilled water for 2 weeks. Urine NOX and 8-iso-PGF2alpha were determined after 2 weeks.


RESULTS
L-NMMA reduced both urine NOX and 8-iso-PGF2alpha levels in a dose-dependent fashion in NZB/W and MRL/lpr mice. Urine NOX and 8-iso-PGF2alpha levels were highly correlated. Both specific (L-NIL) and nonspecific (L-NMMA) iNOS inhibition reduced urine NOX and 8-iso-PGF2alpha levels in both models of murine PLN.


CONCLUSIONS
These findings suggest that iNOS activity is a major source of reactive oxidant stress in these models of murine PLN. Future studies will address the pathogenic role of reactive oxygen stress in PLN.
Njoku
Chinedu J
CJ
Department of Medicine, Division of Rheumatology, Medical University of South Carolina, Charleston, USA.
Patrick
Kennerly S
KS
Ruiz
Philip
P
Oates
Jim C
JC
D000818:Animals; D015415:Biomarkers / Q000652:urine; D015237:Dinoprost / Q000031:analogs & derivatives / Q000652:urine; D004791:Enzyme Inhibitors / Q000008:administration & dosage / Q000494:pharmacology*; D005260:Female; D008181:Lupus Nephritis / Q000188:drug therapy* / Q000378:metabolism; D008239:Lysine / Q000031:analogs & derivatives / Q000494:pharmacology; D051379:Mice; D008814:Mice, Inbred NZB; D009566:Nitrates / Q000652:urine; D052247:Nitric Oxide Synthase Type II / Q000037:antagonists & inhibitors*; D009573:Nitrites / Q000652:urine; D018384:Oxidative Stress / Q000187:drug effects*; D019323:omega-N-Methylarginine / Q000008:administration & dosage / Q000494:pharmacology
D015415:Biomarkers; D004791:Enzyme Inhibitors; C090663:N(6)-(1-iminoethyl)lysine; D009566:Nitrates; D009573:Nitrites; C075750:8-epi-prostaglandin F2alpha; D019323:omega-N-Methylarginine; D015237:Dinoprost; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; D008239:Lysine