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Effect of psychosine on inducible nitric-oxide synthase expression under different culture conditions: implications for Krabbe disease.
[krabbe disease]
Krabbe
disease
is
a
neuro-
inflammatory
disorder
in
which
galactosyl
sphingosine
(
psychosine
)
accumulates
in
nervous
tissues
.
Despite
some
leads
in
elucidating
the
mechanism
of
psychosine
action
on
different
cells
of
brain
,
there
still
remain
gaps
in
the
knowledge
and
mechanisms
behind
events
in
the
pathogenic
cascade
of
Krabbe
disease
.
Inflammation
in
the
brain
in
Krabbe
disorder
is
an
important
factor
in
neural
damage
.
This
study
was
undertaken
to
access
the
role
of
psychosine
in
the
regulation
of
nitric
oxide
(
NO
)
and
inducible
nitric
oxide
synthase
(
iNOS
)
,
i
.
e
.
,
inflammatory
markers
,
under
two
different
conditions
viz
.
,
using
a
single
cell
line
and
using
primary
mixed
glial
cells
.
BV
2
murine
microglial
cells
and
murine
primary
mixed
glial
cells
were
used
during
this
study
.
The
cell
lines
,
after
12
hr
serum
starvation
,
were
treated
with
lipopolysaccharide
(
LPS
)
at
25
ng
/
ml
in
the
absence
or
presence
of
increasing
concentrations
of
psychosine
(
5
,
10
and
15
microM
)
.
Formation
of
NO
was
estimated
using
Greiss
reagent
,
and
expression
of
iNOS
was
done
by
SDS
-page
followed
by
western
blotting
using
anti-i
NOS
antibody
.
I
n
BV
2
cells
it
was
found
that
LPS
(
25
ng
/
ml
)
treatment
,
induced
production
of
NO
,
but
the
LPS
induced
NO
production
was
inhibited
when
LPS
was
used
in
combination
with
psychosine
.
This
result
was
corroborated
by
parallel
trend
seen
in
iNOS
expression
under
same
conditions
.
Contrary
to
this
,
LPS
(
25
ng
/
ml
)
induced
production
of
NO
in
primary
mixed
glial
cells
was
dose
dependently
enhanced
when
LPS
was
used
in
combination
with
psychosine
(
5
,
10
and
15
microM
)
.
This
result
was
also
corroborated
by
iNOS
expression
under
same
experimental
conditions
.
This
study
suggests
that
in
-vitro
data
obtained
using
individual
cell
lines
may
not
reflect
the
actual
complex
intricacies
involved
in
development
of
Krabbe
brain
pathology
.
And
that
the
effect
of
psychosine
in
Krabbe
brain
may
be
modulated
by
presence
of
LPS
or
other
pro-
inflammatory
stimuli
in
the
brain
of
these
children
,
e
.
g
.
,
after
an
infection
.
Diseases
Validation
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"e"
symptom
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