Detoxification of H2O2 by cultured rabbit lens epithelial cells: Participation of the glutathione redox cycle

Giblin, Frank J.; McCready, Janet P.; Reddan, John R.; Dziedzic, Dorothy C.; Reddy, Venkat N.

doi:10.1016/0014-4835(85)90128-9

Cited by 57 publications

(33 citation statements)

References 28 publications

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“…The determination of GSH levels in both HLE-B3 and BLECs treated overnight (20 h) with or without 1 mM L-BSO was performed as previously described (Giblin et al, 1985). Briefly, the culture medium was removed and a 0.2 ml volume of ice cold 0.05 M EDTA pH 4.0-4.5 was added to the monolayer and the cells were gently scraped off with a cell scraper (Becton Dickinsin Labware, Franklin Lakes, NJ).…”

Section: Measurement Of Glutathione (Gsh)mentioning

confidence: 99%

17β-estradiol stimulates MAPK signaling pathway in human lens epithelial cell cultures preventing collapse of mitochondrial membrane potential during acute oxidative stress

Moor¹,

Flynn²,

Gottipati³

et al. 2005

Mitochondrion

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Abstract17β-estradiol (17β-E 2 ) protects against H 2 O 2 -mediated depletion of intracellular ATP and lessens the degree of depolarization of mitochondrial membrane potential (Δψ m ) in cultured lens epithelial cells consequential to oxidative insult. We now report that 17β-E 2 acts as a positive regulator of the survival signal transduction pathway, MAPK which, in turn, acts to stabilize Δψ m , in effect, attenuating the extent of depolarization of mitochondrial membrane potential in the face of acute oxidative stress. The SV-40 viral transformed human cell line, HLE-B3 was treated with 17β-E 2 over a time course of 60 min and phosphorylation of ERK1/2 was analyzed by Western blot. ERK1/2 was phosphorylated within 5-15 min in the presence of 17β-E 2 . Cell cultures were exposed to the MEK1/2 inhibitor, UO126, subsequent to H 2 O 2 ±17β-E 2 treatment and the Δψ m examined using JC-1, a potentiometric dye which serves as an indicator for the state of mitochondrial membrane potential. UO126 treatment attenuated ERK1/2 phosphorylation irrespective of whether estradiol was administered. Mitochondrial membrane depolarization resulting from H 2 O 2 stress was substantially greater in the presence of UO126. The greater the extent of depolarization, the less effective 17β-E 2 treatment was in checking mitochondrial membrane depolarization, indicating that the relative degree of ERK phosphorylation influences mitochondrial stability with oxidative insult. The data support a positive correlation between 17β-E 2 stimulation of ERK1/2 phosphorylation and mitochondrial stabilization that would otherwise cause a complete collapse of Δψ m .

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Section: Measurement Of Glutathione (Gsh)mentioning

confidence: 99%

17β-estradiol stimulates MAPK signaling pathway in human lens epithelial cell cultures preventing collapse of mitochondrial membrane potential during acute oxidative stress

Moor¹,

Flynn²,

Gottipati³

et al. 2005

Mitochondrion

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“…In different biological systems, the GSH redox cycle plays an important role in protecting cells against oxidative damage (93)(94)(95). Generally, GSH is present in millimolar concentrations in cytosol and the nucleus, whereas its concentration is low in blood serum and other biological fluids (14,87).…”

Section: Seminal Fluid and Ros Damagementioning

confidence: 99%

Lipoperoxidation damage of spermatozoa polyunsaturated fatty acids PUFA scavenger mechanisms and possible scavenger therapies

Lenzi¹

2000

Front Biosci

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The lipid metabolism in sperm cells is important both as one of the main sources for energy production and for cell structure. The double leaflets of the membrane should be considered not simply as a passive lipid film, but as a very specialized structure. The complete maturation of the sperm cell membrane is attained after testicular lipid biosynthetic processes and after passage through the epididymis. A special composition of membrane phospholipids, rich in polyunsaturated fatty acids (PUFA), and the different composition of sperm and immature germ cell membrane are described and discussed.Testis germ cells as well as epididymal maturing spermatozoa are endowed with enzymatic and nonenzymatic scavenger systems to prevent lipoperoxidative damage. Catalase, superoxide dismutase and GSHdependent oxidoreductases are present in variable amounts in the different developmental stages. Phospholipid hydroperoxide GSH peroxidase (PHGPx) activity and alpha tochopherol of epididymal spermatozoa are considered in detail. Their distribution and roles in caput and cauda epididymal sperm cells are discussed.Seminal plasma also has a highly specialized scavenger system that defends the sperm membrane against lipoperoxidation and the degree of PUFA insaturation acts to achieve the same goal. Systemic predisposition and a number of pathologies can lead to an anti-oxidant/prooxidant disequilibrium. Scavengers, such as GSH, can be used to treat these cases as they can restore the physiological constitution of PUFA in the cell membrane. The results of GSH therapy are presented and discussed. PHOSPHOLIPIDS OF THE SPERM PLASMA MEMBRANE: THEIR PHYSIOLOGY AND ROLE Preliminary remarksSperm membranes play a very active role in sperm fertilization capacity and in sperm-oocyte cross talk, and its biochemical constitution is one of the main field of interest in the study of sperm physiology and pathology. Spermatozoa are polarized cells with structurally and functionally distinct domains. The two leaflets in the membrane of the cap region, overlying the acrosomal vesicle are the domains sensitive to the capacitation stimuli (1). When the various steps of capacitation have induced an increase in the fluidity of the cap region, a fusogenic process, which possesses already the structural premise, starts between this membrane and the outer acrosomal vesicle membrane. The final event consists in the formation of pores that allow a dispersion of the acrosomal enzymes acrosine and hyaluronidase. A

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“…Peroxide is known to oxidize cysteine and methionine efficiently (11,12). H 2 O 2 may be derived from the aqueous humor which bathes the anterior segment of the lens (13) and is the primary source of nutrients and oxygen, although the lens is remarkable in its ability to detoxify external peroxide (14,15). Since the human lens grows throughout life by the addition of fiber cells, and since the crystallins in the lens nucleus undergo little turnover (16), the proteins in the central region of the lens are as old as the individual.…”

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confidence: 99%

The Hydroxyl Radical in Lens Nuclear Cataractogenesis

Dean

Southan³

et al. 1998

Journal of Biological Chemistry

161

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Cataract is the major cause of blindness; the most common form is age-related, or senile, cataract. The reasons for the development of cataract are unknown. Here we demonstrate that nuclear cataract is associated with the extensive hydroxylation of protein-bound amino acid residues, which increases with the development of cataract by up to 15-fold in the case of DOPA. The relative abundance of the oxidized amino acids in lens protein (assessed per parent amino acid) is DOPA > o-and m-tyrosine > 3-hydroxyvaline, 5-hydroxyleucine > dityrosine. Nigrescent cataracts, in which the normally transparent lens becomes black and opaque, contain the highest level of hydroxylated amino acids yet observed in a biological tissue: for example, per 1000 parent amino acid residues, DOPA, 15; 3-hydroxyvaline, 0.3; compared with dityrosine, 0.05. The products include representatives of the hydroperoxide and DOPA pathways of protein oxidation, which can give rise to secondary reactive species, radical and otherwise. The observed relative abundance corresponds closely with that of products of hydroxyl radical or metal-dependent oxidation of isolated proteins, and not with the patterns resulting from hypochlorite or tyrosyl-radical oxidation. Although very little light in the 300 -400-nm range passes the cornea and the filter compounds of the eye, we nevertheless also demonstrate that photoxidation of lens proteins with light of 310 nm, the part of the spectrum in which protein aromatic residues have residual absorbance, does not give rise to the hydroxylated aliphatic amino acids. Thus the post-translational modification of crystallins by hydroxyl radicals/Fenton systems seems to dominate their in vivo oxidation, and it could explain the known features of such nuclear cataractogenesis.

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Detoxification of H2O2 by cultured rabbit lens epithelial cells: Participation of the glutathione redox cycle

Cited by 57 publications

References 28 publications

17β-estradiol stimulates MAPK signaling pathway in human lens epithelial cell cultures preventing collapse of mitochondrial membrane potential during acute oxidative stress

17β-estradiol stimulates MAPK signaling pathway in human lens epithelial cell cultures preventing collapse of mitochondrial membrane potential during acute oxidative stress

Lipoperoxidation damage of spermatozoa polyunsaturated fatty acids PUFA scavenger mechanisms and possible scavenger therapies

The Hydroxyl Radical in Lens Nuclear Cataractogenesis

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