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The antioxidant defense system of isolated guinea pig Leydig cells

The antioxidant defense system of isolated guinea pig Leydig cells - Cell Biology and Cell Culture

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Old 11-29-2008, 04:05 PM
tocopherol
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Default The antioxidant defense system of isolated guinea pig Leydig cells



The antioxidant defense system of isolated guinea pig Leydig cells

by Mark A. Kukucka and Hara P. Misra
Department of Biomedical Sciences
Virginia-Maryland Regional College of Veterinary Medicine
Virginia Polytechnic Institute & State University, Blacksburg 24061


ABSTRACT

Utilization of highly enriched preparations of steroidogenic Leydig
cells
have proven invaluable for studying the direct effects of various
hormones
and agents on Leydig cell function in vitro. However, recent work
indicates
that isolated Leydig cells are often subjected to oxygen (O2) toxicity
when
cultured at ambient (19%) oxygen concentrations. Because
intracellular
antioxidants play an important role in protecting cells against
oxygen
toxicity, we have investigated the intracellular antioxidant defense
system
of isolated Leydig cells. The cellular levels of several antioxidants
including catalase, glucose-6-phosphate dehydrogenase (G-6-PDH),
superoxide
dismutase (SOD) of the Cu/Zn & Mn variety, glutathione peroxidase,
glutathione reductase and total glutathione were quantitated using
enriched
populations of Leydig cells isolated from adult male guinea pig
testes.
Compared to whole testicular homogenates, Leydig cells contained
significantly (P < 0.01) less G-6-PDH, total SOD, glutathione
reductase and
total glutathione, but significantly (P < 0.001) more glutathione
peroxidase. Compared to hepatic values previously reported in the
guinea
pig, Leydig cells contain nearly 400 times less catalase, about 14
times
less glutathione peroxidase and almost 11 times less glutathione
reductase.
Since G-6-PDH and glutathione reductase are both necessary to
regenerate
reduced glutathione (GSH) which couples with glutathione peroxidase
to
breakdown hydrogen peroxide (H2O2) under normal conditions, it is
plausible
that the oxygen toxicity observed in isolated Leydig cells is due to
the
intracellular accumulation of H2O2. Using the dichlorofluorescin
diacetate
(DCF-DA) assay, we found that Leydig cells incubated in the presence
of 19%
O2 produced significantly (P<0.001) higher levels of H2O2 with time
in
culture compared to Leydig cells maintained at 3% O2. These results
support
the hypothesis that the increased susceptibility of isolated Leydig
cells to
oxygen toxicity may be due, in part, to decreased amounts of certain
antioxidant defenses and an increased production of the reactive
oxygen
species H2O2.


CITATION
Kukucka, Mark A. and Hara P. Misra. 1993. The antioxidant defense
system of
isolated guinea pig Leydig cells. Mol Cell Biochem 126(1):1-7.


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