Tags

Type your tag names separated by a space and hit enter

NAD(P)H oxidase and uncoupled nitric oxide synthase are major sources of glomerular superoxide in rats with experimental diabetic nephropathy.
Am J Physiol Renal Physiol. 2005 Jun; 288(6):F1144-52.AJ

Abstract

Increased production of reactive oxygen species (ROS) in diabetes may be a common pathway linking diverse pathogenic mechanisms of diabetic vascular complications, including nephropathy. Assessment of the oxidative stress production pathway is therefore important for the prediction and prevention of diabetic complications. However, ROS production mechanisms remain unclear in diabetic glomeruli. To identify the source and determine the mechanisms of ROS production in the diabetic kidney, diabetes was induced with streptozotocin in rats. After 6 wk, glomerular ROS production had increased in the streptozotocin rat kidney, as assessed by dihydroethidium-derived chemiluminescence. ROS production was increased by the addition of NADH or L-arginine and was partially reduced by the addition of diphenylene iodonium or N(G)-nitro-L-arginine methyl ester, identifying NAD(P)H oxidase and nitric oxide (NO) synthase (NOS) as ROS sources. The mRNA and protein expression of endothelial NOS (eNOS), as measured by real-time RT-PCR and Western blotting, increased significantly (mRNA level, 1.3-fold; protein level, 1.8-fold). However, the dimeric form of eNOS was decreased in diabetic glomeruli, as measured by low-temperature SDS-PAGE. Production of renal ROS and NO by uncoupled NOS was imaged by confocal laser microscopy after renal perfusion of 2',7'-dichlorofluorescein diacetate (a ROS marker) and diaminorhodamine-4M AM (a NO marker) with L-arginine. Accelerated ROS production and diminished bioavailable NO caused by NOS uncoupling were noted in the diabetic kidney. Administration of tetrahydrobiopterin (BH4), a cofactor for eNOS, reversed the decreased dimeric form of eNOS and glomerular NO production. Our results indicate that NAD(P)H oxidase and uncoupling of eNOS contribute to glomerular ROS production, mediated by the loss of BH4 availability. These mechanisms are potential key targets for therapeutic interventions.

Authors+Show Affiliations

Satoh M
Div. of Nephrology, Dept. of Internal Medicine, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama 701-0192, Japan. minosatou-npr@umin.ac.jp
Fujimoto S
No affiliation info available
Haruna Y
No affiliation info available
Arakawa S
No affiliation info available
Horike H
No affiliation info available
Komai N
No affiliation info available
Sasaki T
No affiliation info available
Tsujioka K
No affiliation info available
Makino H
No affiliation info available
Kashihara N
No affiliation info available

MeSH

AnimalsAntioxidantsBiopterinDiabetic NephropathiesGene Expression Regulation, EnzymologicKidney GlomerulusMaleNADPH OxidasesNitric OxideNitric Oxide SynthaseNitric Oxide Synthase Type IIIOxidative StressRatsRats, Sprague-DawleySuperoxidesTyrosine

Pub Type(s)

Journal Article

Language

eng

PubMed ID

15687247

Citation

Satoh, Minoru, et al. "NAD(P)H Oxidase and Uncoupled Nitric Oxide Synthase Are Major Sources of Glomerular Superoxide in Rats With Experimental Diabetic Nephropathy." American Journal of Physiology. Renal Physiology, vol. 288, no. 6, 2005, pp. F1144-52.
Satoh M, Fujimoto S, Haruna Y, et al. NAD(P)H oxidase and uncoupled nitric oxide synthase are major sources of glomerular superoxide in rats with experimental diabetic nephropathy. Am J Physiol Renal Physiol. 2005;288(6):F1144-52.
Satoh, M., Fujimoto, S., Haruna, Y., Arakawa, S., Horike, H., Komai, N., Sasaki, T., Tsujioka, K., Makino, H., & Kashihara, N. (2005). NAD(P)H oxidase and uncoupled nitric oxide synthase are major sources of glomerular superoxide in rats with experimental diabetic nephropathy. American Journal of Physiology. Renal Physiology, 288(6), F1144-52.
Satoh M, et al. NAD(P)H Oxidase and Uncoupled Nitric Oxide Synthase Are Major Sources of Glomerular Superoxide in Rats With Experimental Diabetic Nephropathy. Am J Physiol Renal Physiol. 2005;288(6):F1144-52. PubMed PMID: 15687247.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - NAD(P)H oxidase and uncoupled nitric oxide synthase are major sources of glomerular superoxide in rats with experimental diabetic nephropathy. AU - Satoh,Minoru, AU - Fujimoto,Sohachi, AU - Haruna,Yoshisuke, AU - Arakawa,Sayaka, AU - Horike,Hideyuki, AU - Komai,Norio, AU - Sasaki,Tamaki, AU - Tsujioka,Katsuhiko, AU - Makino,Hirofumi, AU - Kashihara,Naoki, Y1 - 2005/02/01/ PY - 2005/2/3/pubmed PY - 2005/6/15/medline PY - 2005/2/3/entrez SP - F1144 EP - 52 JF - American journal of physiology. Renal physiology JO - Am J Physiol Renal Physiol VL - 288 IS - 6 N2 - Increased production of reactive oxygen species (ROS) in diabetes may be a common pathway linking diverse pathogenic mechanisms of diabetic vascular complications, including nephropathy. Assessment of the oxidative stress production pathway is therefore important for the prediction and prevention of diabetic complications. However, ROS production mechanisms remain unclear in diabetic glomeruli. To identify the source and determine the mechanisms of ROS production in the diabetic kidney, diabetes was induced with streptozotocin in rats. After 6 wk, glomerular ROS production had increased in the streptozotocin rat kidney, as assessed by dihydroethidium-derived chemiluminescence. ROS production was increased by the addition of NADH or L-arginine and was partially reduced by the addition of diphenylene iodonium or N(G)-nitro-L-arginine methyl ester, identifying NAD(P)H oxidase and nitric oxide (NO) synthase (NOS) as ROS sources. The mRNA and protein expression of endothelial NOS (eNOS), as measured by real-time RT-PCR and Western blotting, increased significantly (mRNA level, 1.3-fold; protein level, 1.8-fold). However, the dimeric form of eNOS was decreased in diabetic glomeruli, as measured by low-temperature SDS-PAGE. Production of renal ROS and NO by uncoupled NOS was imaged by confocal laser microscopy after renal perfusion of 2',7'-dichlorofluorescein diacetate (a ROS marker) and diaminorhodamine-4M AM (a NO marker) with L-arginine. Accelerated ROS production and diminished bioavailable NO caused by NOS uncoupling were noted in the diabetic kidney. Administration of tetrahydrobiopterin (BH4), a cofactor for eNOS, reversed the decreased dimeric form of eNOS and glomerular NO production. Our results indicate that NAD(P)H oxidase and uncoupling of eNOS contribute to glomerular ROS production, mediated by the loss of BH4 availability. These mechanisms are potential key targets for therapeutic interventions. SN - 1931-857X UR - https://www.unboundmedicine.com/medline/citation/15687247/NAD_P_H_oxidase_and_uncoupled_nitric_oxide_synthase_are_major_sources_of_glomerular_superoxide_in_rats_with_experimental_diabetic_nephropathy_ DB - PRIME DP - Unbound Medicine ER -