International Journal of Molecular Sciences,
Год журнала:
2023,
Номер
24(19), С. 14994 - 14994
Опубликована: Окт. 8, 2023
This
study
investigated
the
role
of
a
pattern
microRNA
(miRNA)
as
possible
mediators
celecoxib
and
prescription-grade
glucosamine
sulfate
(GS)
effects
in
human
osteoarthritis
(OA)
chondrocytes.
Chondrocytes
were
treated
with
(1.85
µM)
GS
(9
µM),
alone
or
combination,
for
24
h,
without
interleukin
(IL)-1β
(10
ng/mL).
Cell
viability
was
determined
using
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide
(MTT)
assay,
apoptosis
reactive
oxygen
species
(ROS)
by
cytometry,
nitric
oxide
(NO)
Griess
method.
Gene
levels
miRNA,
antioxidant
enzymes,
nuclear
factor
erythroid
(NRF)2,
B-cell
lymphoma
(BCL)2
expressions
analyzed
quantitative
real
time
polymerase
chain
reaction
(real
PCR).
Protein
expression
NRF2
BCL2
also
detected
at
immunofluorescence
western
blot.
Celecoxib
GS,
significantly
increased
viability,
reduced
apoptosis,
ROS
NO
production
gene
miR-34a,
-146a,
-181a,
-210,
comparison
to
baseline
IL-1β.
The
transfection
miRNA
specific
inhibitors
counteracted
IL-1β
activity
potentiated
properties
on
oxidant
system,
through
(NF)-κB
regulation.
observed
enhanced
when
drugs
tested
combination.
Our
data
confirmed
synergistic
anti-inflammatory
chondroprotective
suggesting
mediators.
Research Square (Research Square),
Год журнала:
2024,
Номер
unknown
Опубликована: Май 22, 2024
Abstract
Background
and
Aim:
Understanding
the
structural
basis
of
Keap1,
a
central
regulator
Nrf2
pathway
is
crucial
for
cancer
regulation.
The
recent
crystallographic
elucidation
Keap1's
structure
provides
insights
into
its
functional
domains
potential
ligand
binding
sites,
paving
way
targeted
drug-discovery
efforts.
This
study
aims
to
identify
small
molecule
with
high
affinity
against
Keap1
as
modulator
SQSTM1/p62,
function
in
colorectal
(CRC)
cells.
Methods
A
high-throughput
virtual
screening
approach
was
used
screen
ChemBridge
library
protein.
Atomistic
Molecular
Dynamics
(MD)
simulations
were
conducted
using
GROMACS,
along
Gibbs
free
energy
estimations.
HCT116
Caco-2
cells
determine
anti
proliferation.
Flow
cytometry
evaluate
target
inhibition
Results
Identified
CBCB5712809,
exhibited
stable
avid
interactions
key
residues
Keap1.
dynamics
demonstrated
stability
protein-ligand
complex
over
200ns
trajectory.
MM-PBSA
analysis
indicated
favorable
interaction
between
CBCB5712809
suggesting
function.
suppressed
growth
GI50
values
40.07
nM
102.80
respectively.
shows
that
arrested
CRC
G2/M
phase
cell
cycle
downregulated
levels
while
upregulating
SQSTM1/p62
levels.
Conclusion
this
further
experimental
validation
develop
chemotherapeutic
CRC.
International Journal of Molecular Sciences,
Год журнала:
2023,
Номер
24(19), С. 14994 - 14994
Опубликована: Окт. 8, 2023
This
study
investigated
the
role
of
a
pattern
microRNA
(miRNA)
as
possible
mediators
celecoxib
and
prescription-grade
glucosamine
sulfate
(GS)
effects
in
human
osteoarthritis
(OA)
chondrocytes.
Chondrocytes
were
treated
with
(1.85
µM)
GS
(9
µM),
alone
or
combination,
for
24
h,
without
interleukin
(IL)-1β
(10
ng/mL).
Cell
viability
was
determined
using
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide
(MTT)
assay,
apoptosis
reactive
oxygen
species
(ROS)
by
cytometry,
nitric
oxide
(NO)
Griess
method.
Gene
levels
miRNA,
antioxidant
enzymes,
nuclear
factor
erythroid
(NRF)2,
B-cell
lymphoma
(BCL)2
expressions
analyzed
quantitative
real
time
polymerase
chain
reaction
(real
PCR).
Protein
expression
NRF2
BCL2
also
detected
at
immunofluorescence
western
blot.
Celecoxib
GS,
significantly
increased
viability,
reduced
apoptosis,
ROS
NO
production
gene
miR-34a,
-146a,
-181a,
-210,
comparison
to
baseline
IL-1β.
The
transfection
miRNA
specific
inhibitors
counteracted
IL-1β
activity
potentiated
properties
on
oxidant
system,
through
(NF)-κB
regulation.
observed
enhanced
when
drugs
tested
combination.
Our
data
confirmed
synergistic
anti-inflammatory
chondroprotective
suggesting
mediators.
