[8] Chemical depletion of glutathione in vivo

Plummer, John L.; Smith, Brian R.; Sies, Helmut; Bend, John R.

doi:10.1016/s0076-6879(81)77010-1

Cited by 207 publications

(99 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

Section: Wavelength (Nm)mentioning

confidence: 99%

“…1 to staining (8). Nonprotein thiol depletion was induced by incubation of the cell monolayers with 5-10 mM diethylmaleate (DEM) (Sigma Chemical Co., St. Louis, MO) for up to 6 h prior to staining (8). Specific depletion of GSH was accomplished by incubation of cells overnight in 1-10 mM DL-buthionine-S, R-sulfoximine (BSO) (Chemical Dynamics Corp., South Plainfield, NJ).…”

Section: Wavelength (Nm)mentioning

confidence: 99%

See 1 more Smart Citation

A flow cytometric assay for intracellular nonprotein thiols using mercury orange

et al. 1988

View full text Add to dashboard Cite

The level of nonprotein thiols was assayed in individual mammalian cells using flow cytometry. Previous determinations of glutathione (GSH, the most abundant nonprotein thiol in most cells) by flow cytometry were based on UV laser excitation of fluorochromes. Because of several shortcomings of W excitation, an assay for GSH using visible light is of interest. Selective staining of nonprotein thiols with mercury orange (a mercurial compound that binds stoichiometrically to sulfhydry1 groups) was obtained by restricting the staining time. By using various drugs that affect GSH levels and overall thiol levels in cells, it was shown that GSH is the primary thiol group being stained. Thus a quick, specific technique using mercury orange has been developed for the flow cytometric determination of nonprotein thiols and preferentially for GSH in individual mammalian cells. Key terms: Glutathione, mammalian cellsGlutathione (GSH) plays a ubiquitous and important role in many physiological processes, including free radical scavenging, xenobiotic detoxification, amino acid transport, thermosensitivity and thermotolerance, protection against ionizing radiation and chemical carcinogenesis, and maintenance of cellular structures (for a review, 6). A number of fluorochromes have been tested for thiol staining and GSH determination by flow cytometry (3,7,9,10), but most of them are based on W excitation measurements. Unfortunately, many flow cytometers utilize lasers that operate exclusively in the visible spectrum. Moreover, the use of W excitation dramatically shortens the lifetime of the laser tube. Thus a n assay for GSH in the visible light range is of considerable practical interest. To our knowledge, only the fluorochrome fluorescein-5-maleimide (F5M) has been applied to the determination of GSH by fluorescent flow cytometry at 488-nm excitation (3). Nevertheless, its use was limited because of its poor penetration into cells and its low relative fluorescent yield when compared with other fluorochromes.In 1975, Asghar et al.(1) reported a quick and easy method for staining GSH in frozen tissue sections, based on the reaction with mercury orange in toluene. The rate of reaction of mercury orange and GSH was found to be considerably faster than with protein thiols. After a short incubation time, more than 75-80% of the GSH had reacted, while at least a n 8-h period was required for the mercury orange to react with protein SH groups.Recently, Larrauri et al. (5) published a technical note on a modification of Asghar's procedure and on the effect of different solvents on the staining efficiency. From the works of Asghar et al. and Larrauri et al., it seemed that mercury orange could be used for GSH determination by flow cytometry, since the reaction product emitted intense red fluorescence when excited in the blue light region, and reactions were quantitative when using appropriate solvents and incubation times, In this paper, we demonstrate that mercury orange can be used for flow cytometric determination of GSH using the 4...

show abstract

Section: Wavelength (Nm)mentioning

confidence: 99%

Section: Wavelength (Nm)mentioning

confidence: 99%

A flow cytometric assay for intracellular nonprotein thiols using mercury orange

et al. 1988

View full text Add to dashboard Cite

show abstract

“…Recently the dependence of ascorbate synthesis on glycogenolysis has been reported [6]. However, glycogenolysis is also induced by agents, which provoke GSH deficiency [7][8][9].…”

Section: Introductionmentioning

confidence: 99%

Glutathione depletion induces glycogenolysis dependent ascorbate synthesis in isolated murine hepatocytes

et al. 1996

View full text Add to dashboard Cite

The relationship between glutathione deficiency, glycogen metabolism and ascorbate synthesis was investigated in isolated murine hepatocytes. Glutathione deficiency caused by various agents increased ascorbate synthesis with a stimulation of glycogen breakdown. Increased ascorbate synthesis from UDP-glucose or gulonolactone could not be further affected by glutathione depletion. Fructose prevented the stimulated glycogenolysis and ascorbate synthesis caused by glutathione consumption. Reduction of oxidised glutathione by dithiothreitol decreased the elevated glycogenolysis and ascorbate synthesis in diamide or menadione treated hepatocytes. Our results suggest that a change in GSI-I/GSSG ratio seems to be a sufficient precondition of altering glycogenolysis and a consequent ascorbate synthesis.

show abstract

“…Nitrosothiols have been shown to release NO and to stimulate guanylate cyclase [35,[52][53][54]. Moreover, pretreatment with diethyl maleate, an agent known to deplete tissue GSH [55], reduced both relaxation and formation of NO upon exposure to PN. It was concluded that PN reacts with arterial smooth muscle to generate NO that leads to initial relaxation of vascular smooth muscle, but that the decomposition products of PN also contribute to the prolonged response by the generation of a nitrosothiol-like compound that continues to release NO over prolonged periods [21].…”

Section: Vasorelaxant Properties Of Peroxyni-tritementioning

confidence: 99%

Potential Benefits of Peroxynitrite

Nossaman

Kadowitz²

2008

TOPHARMJ

View full text Add to dashboard Cite

Peroxynitrite (PN) is generated by the reaction of nitric oxide (NO) and superoxide in one of the most rapid reactions in biology. Studies have reported that PN is a cytotoxic molecule that contributes to vascular injury in a number of disease states. However, it has become apparent that PN has beneficial effects including vasodilation, inhibition of platelet aggregation, inhibition of inflammatory cell adhesion, and protection against ischemia/reperfusion injury in the heart. It is our hypothesis that PN may serve to inactivate superoxide and prolong the actions of NO in the circulation. This manuscript reviews the beneficial effects of PN in the cardiovascular system. Keywords:Nitric oxide/*metabolism, nitric oxide synthase/metabolism/physiology, peroxynitrous acid/metabolism, reactive nitrogen species/metabolism, reactive oxygen species/metabolism, endothelium, vascular/drug effects/metabolism, muscle, smooth, vascular/drug effects/*metabolism, vasodilator agents/adverse effects, oxidative stress/*physiology, vasodilation/drug effects.

show abstract

[8] Chemical depletion of glutathione in vivo

Cited by 207 publications

References 49 publications

A flow cytometric assay for intracellular nonprotein thiols using mercury orange

A flow cytometric assay for intracellular nonprotein thiols using mercury orange

Glutathione depletion induces glycogenolysis dependent ascorbate synthesis in isolated murine hepatocytes

Potential Benefits of Peroxynitrite

Contact Info

Product

Resources

About