title: Indomethacin inhibits activation of endothelial nitric oxide synthase in the rat kidney: possible role of this effect in the pathogenesis of indomethacin-induced renal damage.
abstract: The clinical use of non-steroidal anti-inflammatory drugs (NSAIDs) is often associated with adverse effects in the kidney. Indomethacin, an NSAID that has been shown to induce oxidative stress in the kidney, was used to study the pathogenesis of renal damage induced by the drug in a rat model. Experimental animals were given indomethacin (20mg/kg) by oral gavage, sacrificed 1, 12 or 24h (h) later and the kidneys studied. Evidence of glomerular and tubular damage in the kidney was found in response to the drug. Renal tissue nitrite levels, a surrogate marker of nitric oxide (NO) synthesis, were significantly decreased at 12 and 24h. Indomethacin did not affect protein and mRNA levels of endothelial nitric oxide synthase (eNOS) or inducible NOS (iNOS). However, it significantly reduced the ratio of dimeric (active) to monomeric (inactive) eNOS in the kidney, 12 and 24h after drug administration. This was associated with reductions in heme content, both in renal tissue and in NOS. Heme oxygenase 1 (HO-1) mRNA (at 1 and 12h), protein (at 12 and 24h) and activity (at 1, 12 and 24h) were elevated in response to indomethacin. Nuclear translocation of Nrf2 (at 12h) and p38 MAPK signaling (at 12h and 24h), both of which are known to induce HO-1, also occurred in response to the drug. In summary, our results show that indomethacin reduced levels of activated eNOS in the kidney. This effect is possibly mediated by heme depletion, secondary to HO-1 induction that occurred downstream of Nrf2 and p38 MAPK signaling. We postulate that reduced renal eNOS activity may result in decreased NO levels, and hence reduced renal perfusion, leading to glomerular and tubular injury with subsequent renal damage.
authors.affiliation: Department of Biochemistry, Christian Medical College, Vellore 632002, India. Electronic address: arumugam_sn@cmcvellore.ac.in.
authors.forename: Arumugam Suriyam
authors.initials: AS
authors.lastname: Nagappan
authors.affiliation: Department of Biochemistry, Christian Medical College, Vellore 632002, India. Electronic address: joevarghese@cmcvellore.ac.in.
authors.forename: Joe
authors.initials: J
authors.lastname: Varghese
authors.affiliation: Department of Biochemistry, Christian Medical College, Vellore 632002, India. Electronic address: gautham.pranesh@gmail.com.
authors.forename: Gautham Tumkur
authors.initials: GT
authors.lastname: Pranesh
authors.affiliation: Department of Biostatistics, Christian Medical College, Vellore 632002, India. Electronic address: visali_pv@hotmail.com.
authors.forename: Visalakshi
authors.initials: V
authors.lastname: Jeyaseelan
authors.affiliation: Department of Biochemistry, Christian Medical College, Vellore 632002, India. Electronic address: mjacob@cmcvellore.ac.in.
authors.forename: Molly
authors.initials: M
authors.lastname: Jacob
mesh_terms: D000284:Administration, Oral; D000818:Animals; D015153:Blotting, Western; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D042783:Endothelial Cells / Q000201:enzymology; D051547:Heme Oxygenase-1 / Q000378:metabolism; D007213:Indomethacin / Q000494:pharmacology*; D007668:Kidney / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D007674:Kidney Diseases / Q000139:chemically induced* / Q000201:enzymology / Q000503:physiopathology; D016014:Linear Models; D008297:Male; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
chemical_list: D016861:Cyclooxygenase Inhibitors; D019001:Nitric Oxide Synthase; D051547:Heme Oxygenase-1; D007213:Indomethacin
keywords: Heme oxygenase-1; Indomethacin; Kidney; NSAID; Nitric oxide synthase type III; Oxidative stress