Chemiluminescent Detection of Oxidants in Vascular Tissue

Tarpey, Margaret M.; White, C. Roger; Suarez, Edward C.; Richardson, Gloria R.; Radí, Rafael; Freeman, Bruce Α.

doi:10.1161/01.res.84.10.1203

Cited by 144 publications

(39 citation statements)

References 77 publications

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“…NO also caused an increase in coelenterazine chemiluminescence in the control cells, but this chemiluminescence was not further increased by chloramphenicol treatment. The probe used here measures both superoxide and peroxynitrite and these cannot be distinguished using these protocols (23).…”

Section: Discussionmentioning

confidence: 99%

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Inhibition of mitochondrial protein synthesis results in increased endothelial cell susceptibility to nitric oxide-induced apoptosis

Ramachandran

Moellering

Ceaser

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

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Mutations in mitochondrial DNA, affecting the activity of respiratory complexes, have been implicated in many chronic degenerative diseases. Mitochondrial proteins coded for by both the mitochondrial and nuclear genes are known to have important signaling roles in apoptosis. However, the impact of the inhibition of mitochondrial protein synthesis on apoptosis is largely unknown. This inhibition is particularly important in NO-dependent cytotoxicity, which is believed to have a significant mitochondrial component and depend on other factors such as glycolysis. In this study we have examined whether the inhibition of mitochondrial protein synthesis by chloramphenicol increases the susceptibility of endothelial cells to undergo NO-dependent apoptosis in glucose-free media. Bovine aortic endothelial cells were treated with chloramphenicol, which resulted in a decreased ratio of mitochondrial complex IV to cytochrome c and increased oxidant production in the cell. Inhibition of mitochondrial protein synthesis was associated with a greater susceptibility of the cells to apoptosis induced by NO in glucose-free medium.

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Section: Discussionmentioning

confidence: 99%

“…Chemiluminescence was then monitored for 2 min by using an Autolumat LB953 luminometer (23). The ROS͞RNS production is expressed as relative light units per 10 6 cells.…”

Section: Measurement Of Reactive Oxygen and Nitrogen Speciesmentioning

confidence: 99%

Inhibition of mitochondrial protein synthesis results in increased endothelial cell susceptibility to nitric oxide-induced apoptosis

Ramachandran

Moellering

Ceaser

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

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“…Vessel O 2 ⅐Ϫ production was monitored in aortic segments by coelenterazineenhanced chemiluminescence (34) by using an automated microplate luminescence reader (Lumistar Galaxy, BMG Lab Technologies). Previous studies have shown coelenterazine to reflect both tissue O 2 ⅐Ϫ and ONOO Ϫ production, differentiable by selective use of SOD and NO synthase inhibitors (34). Isolated vessels were equilibrated in Hanks' balanced salt solution (HBSS) for 30 min at 37°C, and coelenterazine (10 M, Calbiochem) was added.…”

Section: Methodsmentioning

confidence: 99%

Oxygen radical inhibition of nitric oxide-dependent vascular function in sickle cell disease

Aslan

Ryan

Adler

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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349

337

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“…No changes in luminescence were observed in the absence of NADPH, and addition of H 2 O 2 to lucigenin in PBS in the absence of cells did not increase chemiluminescence. Some experiments were also performed with coelenterazine (5 M), a structurally distinct luminescent probe which reportedly is not capable of redox cycling via autoxidation (15). Data were analyzed by ANOVA followed by Bonferroni t testing.…”

Section: Methodsmentioning

confidence: 99%

H2O2-induced O⨪2Production by a Non-phagocytic NAD(P)H Oxidase Causes Oxidant Injury

Li¹,

Miller²,

Zhang³

et al. 2001

Journal of Biological Chemistry

239

165

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Non-phagocytic NAD(P)H oxidases have been impliinduced stimulation of NAD(P)H oxidase activity in nonphagocytic cell types could represent a mechanism by which oxidant-mediated injury is amplified in vivo and identify these NAD(P)H oxidase enzymes as therapeutic targets for the amelioration of the biological effects of chronic oxidant stress. EXPERIMENTAL PROCEDURESCell Culture-SMC were prepared from 250 to 300-g male HarlanSprague Dawley rats as previously described (10, 11). Aortic adventitial fibroblasts were prepared from C57 BL/6 and from gp91 phox knock-out mice (Jackson Lab, Bar Harbor, ME) as described (12, 13). The cells were grown in minimum essential media supplemented with 10% fetal bovine serum and penicillin (100 IU/ml) and streptomycin (100 g/ml) in a humidified atmosphere containing 5% CO 2 at 37°C. Stocks were subcultured at subconfluence by trypsinization. All experiments were performed on cells between passages 8 and 20 grown to 70% to 95% confluence in 24-well plates, 35-or 60-mm dishes, or 4-well chamber slides.Superoxide Measurement by Enhanced Chemiluminescence-Lucigenin luminescence measurements were carried out using 5 M lucigenin, a concentration which is unlikely to induce redox cycling (14). Cells were grown to subconfluence in 35-mm dishes and growth arrested in serum-free medium for 24 h. After exposure to H 2 O 2 at the indicated concentrations and times, cells were washed twice with PBS, and 5 M lucigenin was added. Immediately afterward, NADPH (0.1 mM) was added, and after dark adaptation, luminescence was measured every 15 s for 5 min in a luminometer (Zylux Corp). Changes in luminescence were expressed as relative luminescent units per 10 5 cells per minute. No changes in luminescence were observed in the absence of NADPH, and addition of H 2 O 2 to lucigenin in PBS in the absence of cells did not increase chemiluminescence. Some experiments were also performed with coelenterazine (5 M), a structurally distinct luminescent probe which reportedly is not capable of redox cycling via autoxidation (15). Data were analyzed by ANOVA followed by Bonferroni t testing.Intracellular O 2 . Determination by Confocal Microscopy-SMC grown on chamber slides were treated as indicated under "Results" and in the figure legends, washed, and incubated with dihydroethidium (HE, 5

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Chemiluminescent Detection of Oxidants in Vascular Tissue

Cited by 144 publications

References 77 publications

Inhibition of mitochondrial protein synthesis results in increased endothelial cell susceptibility to nitric oxide-induced apoptosis

Inhibition of mitochondrial protein synthesis results in increased endothelial cell susceptibility to nitric oxide-induced apoptosis

Oxygen radical inhibition of nitric oxide-dependent vascular function in sickle cell disease

H2O2-induced O⨪2Production by a Non-phagocytic NAD(P)H Oxidase Causes Oxidant Injury

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