Objective: To study the effects of resveratrol in neuronal structures in traumatic brain injury (TBI). Study Design: Thirty rats were categorized as (1) control group (n=10), saline solution administered i.p. for 14 days, (2) TBI group (n=10), trauma induced by weight-drop model on brain, and (3) TBI+Resveratrol group (n=10), 15 minutes after injury the rats were given resveratrol (10 μmoL/kg/i.p.) for 14 days. At the end of the experiment the cerebellum was excised for routine paraffin tissue protocol. Blood samples were tested for serum biochemical markers (MDA, SOD, CAT, and GSH-x). Results: SOD, GPx, and CAT values were lowest in the TBI group. MDA and histological scores of dilations in vessels, inflammation, degeneration in neurons, apoptosis in microglia, ADAMTS8, and GFAP expressions were highest in the TBI group. Sections of the control group showed normal cerebellar histology. The trauma group showed degenerated ganglion layer, pyknotic and apoptotic Purkinje cell nuclei. Vascular thrombus was seen in the substantia alba and substantia grisea. In the Trauma+Resveratrol group, most pa- thologies observed in the TBI group were improved. In the control group, GFAP protein was expressed in granular cells, axons, dendrites, Purkinje cells, and microglia cells. In the trauma group, increased GFAP expression was observed in glial processes, neurons, and Purkinje cells. In the Trauma+Resveratrol group, GFAP was expressed in molecular layer and glial processes. In the control group, ADAMTS-4 activity was observed in granulosa layer, glial cells, and Purkinje cells. In the trauma group, ADAMTS-4 expression was positive in Purkinje cells and glial cells. In the Trauma+ Resveratrol group, ADAMTS-4 was expressed in Purkinje cells, granular cells, and glial cells. Conclusion: GFAP and ADAMTS-4 proteins may be involved in regeneration of damaged astroglial cells and other glial cells, Purkinje cells, and synaptic extensions. We suggest that antioxidative drugs such as resveratrol may be alternative target agents in neurological disease. Keywords: ADAMTS-4, brain, cerebellum, GFAP, rat, resveratrol, traumatic brain injury

1. 193
OBJECTIVE: To study the effects of resveratrol in
neuronal structures in traumatic brain injury (TBI).
STUDY DESIGN: Thirty rats were categorized as (1)
control group (n=10), saline solution administered i.p.
for 14 days, (2) TBI group (n=10), trauma induced by
weight-drop model on brain, and (3) TBI+Resveratrol
group (n=10), 15 minutes after injury the rats were
given resveratrol (10 μmoL/kg/i.p.) for 14 days. At the
end of the experiment the cerebellum was excised for
routine paraffin tissue protocol. Blood samples were
tested for serum biochemical markers (MDA, SOD,
CAT, and GSH-x).
RESULTS: SOD, GPx, and CAT values were lowest
in the TBI group. MDA and histological scores of
dilations in vessels, inflammation, degeneration in neu­
rons, apoptosis in microglia, ADAMTS8, and GFAP
expressions were highest in the TBI group. Sections of
the control group showed normal cerebellar histology.
The trauma group showed degenerated ganglion layer,
pyknotic and apoptotic Purkinje cell nuclei. Vascular
thrombus was seen in the substantia alba and substan­
tia grisea. In the Trauma+Resveratrol group, most pa-
thologies observed in the TBI group were improved.
In the control group, GFAP protein was expressed in
granular cells, axons, dendrites, Purkinje cells, and
microglia cells. In the trauma group, increased GFAP
expression was observed in glial processes, neurons,
and Purkinje cells. In the Trauma+Resveratrol group,
GFAP was expressed in molecular layer and glial pro­
cesses. In the control group, ADAMTS-4 activity was
observed in granulosa layer, glial cells, and Purkinje
cells. In the trauma group, ADAMTS-4 expression was
positive in Purkinje cells and glial cells. In the Trauma+
Resveratrol group, ADAMTS-4 was expressed in Pur­
kinje cells, granular cells, and glial cells.
CONCLUSION: GFAP and ADAMTS-4 proteins may
be involved in regeneration of damaged astroglial cells
and other glial cells, Purkinje cells, and synaptic ex-
tensions. We suggest that antioxidative drugs such as
resveratrol may be alternative target agents in neurol­
ogical disease. (Anal Quant Cytopathol Histpathol
2021;43:193–201)
Keywords: ADAMTS-4, brain, cerebellum, GFAP,
rat, resveratrol, traumatic brain injury.
Traumatic brain injury (TBI) is an important cause
of morbidity and mortality for humans and ani-
mals. Some components of TBI have been report­
ed to be partially attributed to cerebellar damage,
including ataxia, postural imbalance, tremors, im-
pairments in balance and fine motor skills, and
cognitive deficits. Oxidative stress, which occurs
with the mass production of free radicals and
lipid peroxides and reactive oxygen species (ROS),
also plays a role in the formation of neurological
vascular injuries.1 In animal model studies on
traumatic brain injury (TBI), pathophysiological
changes have been observed in the cortex and
thalamus, and the hippocampus has been shown
to cause motor and cognitive impairments. Im-
pairments in motor function, coordination, and cer-
ebellar function have been cited among the com­
mon causes of cellular degeneration TBI.2
Analytical and Quantitative Cytopathology and Histopathology®
0884-6812/21/4304-0193/$18.00/0 © Science Printers and Publishers, Inc.
Analytical and Quantitative Cytopathology and Histopathology®
Effect of Resveratrol on the Changes in the
Cerebellum in Traumatic Brain Injury
Mustafa Nevzat Firidin, M.D.
From the Department of Neurosurgery, Faculty of Medicine, Siirt University, Siirt, Turkey.
Mustafa Nevzat Firidin is Assistant Professor.
Address correspondence to: Mustafa Nevzat Firidin, M.D., Department of Neurosurgery, Faculty of Medicine, Siirt University, Gures
Street, 56100 Siirt, Turkey (firidin2828@gmail.com).
Financial Disclosure: The author has no connection to any companies or products mentioned in this article.

2. Resveratrol (3,4′,5-trihydroxystilbene) is the poly-
phenol found in a variety of plants such as grapes,
plums, and peanuts. It is an antioxidant with an-
ticancer, anti-inflammatory, and cardioprotective
properties.3,4 It has been stated that it inhibits
membrane lipid peroxidation, scavenges free rad­
icals by increasing antioxidant levels, and inhibits
platelet aggregation.5 Glial fibrillar acidic protein
(GFAP) is an intermediate filament protein found
in the cytoplasm of astrocyte cells. It has been
shown that GFAP immunoreactivity is a sensitive
indicator of neuronal damage and its increase is
a predictor of reactive astrocytosis. It has been
noted that the level of GFAP in the blood fluid
increases when cerebral tissue or spinal cord cells
are damaged by trauma or disease.6
ADAMTS proteases are a family of
“thrombospondin-motif cleavers and metallopro­
teinases” consisting of 19 members, and an extra­
cellular matrix macromolecule characterized by a
glycosaminoglycan chain channel into perineuro­
nal networks that encircle the neuron subset and
control axon routing and orientation during CNS
development. After CNS damage, CSPGs are rap­
idly upregulated within the glial scar and can have
both beneficial and detrimental effects. Their con­
tribution to a dense glial scar formation initially
creates a protective barrier to limit the spread of
damage, but also creates a deleterious barrier to
subsequent neuro-pair and neuroplasticity.7
In this study, it was aimed to demonstrate the
antioxidative effect and signal stimuli of resvera-
trol, which is an alternative treatment against
oxidative stress and changes in neuronal structure,
which increase in experimental traumatic brain
injury.
Materials and Methods
Traumatic Brain Injury Model Procedure
Thirty animals were divided into 3 groups of 10
each: (1) control group, (2) TBI group, and (3) TBI+
Resveratrol group. The animals were anesthetized
via an intraperitoneal injection of 5 mg/kg xyla-
zine HCl (Rompun, Bayer HealthCare AG, Ger­
many) and 40 mg/kg ketamine HCl (Ketalar,
Pfizer Inc., USA), after which they were allowed
to breathe spontaneously. For the control group
(n=10), isotonic saline solution was administered
intraperitoneally for 14 days. The TBI group (n=
10) was subjected to the diffuse brain injury model
as described by Marmarou et al.8 Briefly, a trauma
device dropped a constant weight (300 g) from
a specific height (1 m) through a tube, inducing
mild trauma. Following trauma, the rats’ cerebel­
lum tissues were removed. For the TBI+ Resvera­
trol group (n=10), 15 minutes following injury, the
rats were given Resveratrol (10 μmoL/kg) intra­
peritoneally every day for 14 days.
Resveratrol was obtained from SIGMA Chemical
(Pool, Dorset). It was added to the drinking water
of the animals following the graft procedure and
was given at 5 mg/kg/day for 14 days. The drink­
ing solutions were changed twice per week and
protected from light in animal drinking bottles.
At the end of the study the animals were
sacrificed, and cerebellum tissues were removed.
Blood samples were collected from the inferior
vena cava for serum biochemical marker deter­
mination in all groups. In this way, MDA, SOD,
CAT, and GSH-x values were measured biochem­
ically. Additionally, cerebellum tissue taken from
the anterior lobe was extracted, fixed in a 10% for­
malin solution, and embedded in paraffin blocks
for histopathologic examination in all groups.
Sections (5 µm thick) were obtained from paraffin
blocks and stained with hematoxylin and eosin
(H&E).
Malondialdehyde and Glutathione Peroxidase Assays
Malondialdehyde (MDA) levels and glutathione
peroxidase (GSH-Px) activities were determined in
the cerebellum of each rat, and the average values
of each group were calculated. Each cerebellum
sample was prepared as a 10% homogenate (accord­
ing to weight) in 0.9% saline using a homogenizer
on ice. Then, the homogenate was centrifuged at
2000 rpm for 10 minutes, and the supernatant was
collected. MDA levels were determined using the
double heating method of Draper and Hadley.9
MDA is an end product of fatty acid peroxidation
that reacts with thiobarbituric acid (TBA) to form
a colored complex. Briefly, 2.5 mL of TBA solution
(100 g/L) was added to 0.5 mL of homogenate in
a centrifuge tube, and the tubes were placed in
boiling water for 15 minutes. After cooling with
flowing water, the tubes were centrifuged at 1000
rpm for 10 minutes, and 2 mL of the supernatant
was added to 1 mL of TBA solution (6.7 g/L); these
tubes were placed in boiling water for another
15 minutes. After cooling, the amount of TBA-
reactive species was measured at 532 nm, and the
MDA concentration was calculated using the ab-
sorbance coefficient of the MDA-TBA complex.
MDA values were expressed as nanomoles per
194 Analytical and Quantitative Cytopathology and Histopathology®
Firidin

3. gram (nmol/g) of wet tissue. The GSH-Px activity
was measured by the method of Paglia and Val­
entine.10 An enzymatic reaction was initiated by
the addition of hydrogen peroxide (H2O2) to a tube
that contained reduced nicotinamide adenine di-
nucleotide phosphate, reduced glutathione, sodi­
um azide, and glutathione reductase. The change
in absorbance at 340 nm was monitored by spec­
trophotometry. Data were expressed as U/g protein.
Measurement of Superoxide Dismutase Activity
Total superoxide dismutase (SOD) activity was de-
termined with a SOD detection kit (RANSOD kit,
Randox Co., UK) according to the manufacturer’s
instructions. SOD accelerates the conversion of
the toxic superoxide (produced during oxidative
energy processes) to hydrogen peroxide and mo-
lecular oxygen. This method employs xanthine and
xanthine oxidase to generate superoxide radicals
that react with 2-(4-iodophenyl)-3-(4-nitrophenol)-
5-phenyltetrazolium chloride (INT) to form a red
formazan dye. The SOD activity is measured by
the degree of inhibition of this reaction. One unit
of SOD causes 50% inhibition of the rate of reduc-
tion of INT under the assay’s conditions. Absor­
bance measurements were taken at 505 nm, and
SOD levels were determined through a standard
curve and expressed as U/mg protein.11
Measurement of Catalase Activity
Tissue catalase (CAT) activity was assayed spectro­
photometrically by monitoring the decomposition
of H2O2 using the procedure of Aebi.12 Briefly, 0.5
mL of 30 mM H2O2 in 50 mM phosphate buffer
(pH 7.0) was added to 1 mL of tissue supernatant
(diluted 1:10), and the consumption of H2O2 was
followed spectrophotometrically at 240 nm for 2
minutes at 25°C. The molar extinction coefficient
was 43.6 L/mol per cm for H2O2. CAT activity was
expressed as mmol H2O2 consumed/min per mg
tissue protein.
Hematoxylin-Eosin Staining Procedure
After the deparaffinizing procedure of sections
with 2 changes of xylene for 10 minutes each, they
were rehydrated in 2 changes of absolute alcohol,
5 minutes each. After being applied with 95% al-
cohol for 2 minutes and 70% alcohol for 2 minutes,
sections were washed briefly in distilled water.
Then, sections were stained in Harris hematoxylin
solution for 8 minutes, washed in running tap
water for 5 minutes, and differentiated in 1% acid
alcohol for 30 seconds. After bluing in 0.2% am-
monia water for 30 seconds, they were washed
in running tap water for 5 minutes and rinsed in
95% alcohol. Then, they were counterstained in
eosin-phloxine solution for 30 seconds and dehy­
drated through 95% alcohol, 2 changes of abso-
lute alcohol, 5 minutes each. They were cleared in
2 changes of xylene, 5 minutes each and mounted
with xylene-based mounting medium.
Immunohistochemical Staining
Antigen retrieval was done in a microwave (Bosch,
700 watt) for 3 min×90°C. They were subjected to
a heating process in a microwave oven at 700 watts
in a citrate buffer (pH 6) solution for proteolysis.
Sections were washed in 3×5 min PBS and incu­
bated with hydrogen peroxide (K-40677109, Merck,
Germany) for 15 min. Sections were washed in
3×5 min PBS and blocked with Ultra V Block (lot
#PHL150128, Thermo Fisher, USA) for 8 minutes.
After draining, primary antibodies were directly
applied to sections distinctly GFAP (Cat. #ab7260,
Abcam, Cambridge, UK) and ADAMTS-4 mono­
clonal antibodies (Cat. #ab185722, Abcam). Sec-
tions were incubated and left overnight at 4°C.
Sections were washed in 3×5 min PBS and then
incubated with biotinylated secondary antibody for
20 minutes. After washing with PBS, streptavidin
peroxidase was applied to sections for 15 minutes.
Sections were washed in 3×5 min PBS and DAB
(lot #HD36221, Thermo Fisher, USA) and were ap-
plied to sections up to 10 minutes. Slides show­
ing reaction were stopped in PBS. Counterstaining
was done with Harris hematoxylin for 45 seconds,
dehydrated through ascending alcohol series, and
cleared in xylene. Slides were mounted with Entel­
lan and examined under light microscope (Zeiss,
Germany).13
Statistical Analysis
The data were recorded as arithmetic mean±stan­
dard deviation with mean rank value. Statistical
analysis was done using the IBM SPSS 25.0 soft-
ware (IBM, Armonk, New York, USA). Kruskal-
Wallis test was used for multiple comparisons.
For within-group comparisons the Mann-Whitney
U test was used. P<0.05 was used as the signifi-
cance level.
Results
Statistical analysis of the biochemical results is
shown in Table I. SOD, GPx, and CAT values
Volume 43, Number 4/August 2021 195
Resveratrol in Traumatic Brain Injury

4. were decreased in the TBI group as compared to
the control and TBI+Resveratrol groups, and the
decrease was statistically significant. MDA value,
dilation in vessels, inflammation, degeneration in
neurons, apoptosis in microglia, and ADAMTS8
and GFAP expressions were increased in the TBI
group as compared to the control and TBI+Res­
veratrol groups, and the increase was statistical-
ly significant. A graphical illustration of Table I is
shown in Figures 1–3.
Histopathologic Examination
In the histological sections of the control group
sections, the nuclei of the granular cells and Pur-
196 Analytical and Quantitative Cytopathology and Histopathology®
Firidin
Table I Biochemical, Histopathological, and Immunohistochemical Scores of the Control, Traumatic Brain Injury (TBI), and
TBI+Resveratrol Groups
Kruskal-Wallis
Mann-Whitney
Mean H test U test
Parameter Group N Mean±SD rank p Value (p<0.05)
SOD (1) Control 10 38.86±0.83 40.19 40.998 (2)(3)
p=0.001
(2) TBI 10 26.47±0.77 8.50 (1)(3)
(3) TBI+Resveratrol 10 32.56±1.93 24.81 (1)(2)
GPx (1) Control 10 346.97±0.75 40.50 41.796 (2)(3)
p=0.001
(2) TBI 10 184.91±0.64 8.50 (1)(3)
(3) TBI+Resveratrol 10 322.65±0.86 24.50 (1)(2)
CAT (1) Control 10 2.57±0.01 40.50 42.155 (2)(3)
p=0.001
(2) TBI 10 1.64±0.01 8.50 (1)(3)
(3) TBI+Resveratrol 10 2.14±0.01 24.50 (1)(2)
MDA (1) Control 10 5.04±0.07 8.50 41.814 (2)(3)
p=0.001
(2) TBI 10 11.01±0.61 40.50 (1)(3)
(3) TBI+Resveratrol 10 8.42±0.61 24.50 (1)(2)
Dilation (1) Control 10 0.5±0.53 6.25 25.401 (2)(3)
p=0.001
(2) TBI 10 3.2±0.42 25.50 (1)(3)
(3) TBI+Resveratrol 10 1.7±0.48 14.75 (1)(2)
Inflammation (1) Control 10 0.6±0.52 6.40 24.976 (2)(3)
p=0.001
(2) TBI 10 3.4±0.52 25.50 (1)(3)
(3) TBI+Resveratrol 10 1.7±0.48 14.60 (1)(2)
Degenerated neurons (1) Control 10 0.7±0.48 6.20 21.789 (2)(3)
p=0.001
(2) TBI 10 3.0±0.67 23.90 (1)(3)
(3) TBI+Resveratrol 10 2.0±0.67 16.40 (1)(2)
Apoptosis in microglia (1) Control 10 0.3±0.48 5.80 25.400 (2)(3)
p=0.001
(2) TBI 10 3.4±0.52 25.20 (1)(3)
(3) TBI+Resveratrol 10 1.9±0.57 15.50 (1)(2)
ADAMTS8 expression (1) Control 10 1.6±0.52 7.30 20.436 (2)(3)
p=0.001
(2) TBI 10 3.2±0.42 23.90 (1)(3)
(3) TBI+Resveratrol 10 2.4±0.52 15.30 (1)(2)
GFAP expression (1) Control 10 2.2±0.79 12.40 14.126 (2)
p=0.001
(2) TBI 10 3.3±0.48 23.40 (1)(3)
(3) TBI+Resveratrol 10 2.1±0.57 10.70 (2)

5. kinje cells were rich in chromatin, and the glomus
part of the granular area appeared to be acido­
philic. No pathological changes were found in
the axonal and dendritic extensions and blood
vessels towards the molecular layer (Figure 4A).
In the trauma group, degeneration and separation
of the ganglionic layer, pyknosis, and apoptosis
were observed in the nuclei of the Purkinje cells.
Thrombus and large-diameter vacuolar structures
were seen in the blood vessels in the substantia
alba and substantia grisea (Figure 4B). In the Trau­
ma+Resveratrol group, there was a decrease in
the Purkinje degenerative appearance, an increase
in chromatin density in the cells of the granular
layer, and an increase in the acidophilic area in the
glomus areas. While no changes were observed in
the glial cells and nerve extensions in substantia
alba, substantia grisea, and also in the blood ves­
sels, a decrease was observed in the vacuolar struc­
tures (Figure 4C).
Immunohistochemical Examination
In the control group sections, GFAP expression
was positive in cells in the granular layer and in
the axons and dendritic extensions, while GFAP
expression was moderate in Purkinje cells and
showed weak expression in microglia cells (Figure
5A). In the Trauma group, increased GFAP expres­
sion was observed with thickening of the dilated
blood vessels in the molecular layer of the glial
footprints. GFAP expression was positive in neu­
ron and glial cells in the granular and gangliosum
layers (Figure 5B). In the Trauma+Resveratrol
group, an increase in the expression of GFAP in
the extensions of the molecular layer and in the
synaptic areas, and regulation in the GFAP expres­
sion in the glial footprints in the substantia alba
region, were observed (Figure 5C). In the control
group, ADAMTS-4 expression was positive in cells
in the granulosa layer, while weak ADAMTS-4
expression was observed in glial cells and Purkinje
cells. In the trauma group, ADAMTS-4 expression
was positive in Purkinje cells, glial cells, and extra­
cellular matrix in the glomus area. In the Trauma+
Resveratrol group, ADAMTS-4 expression was seen
in Purkinje cells, some of the granular cells, and
prominently in glial cells.
Discussion
Traumatic brain injury has been reported to be
among the changes it has made in the cerebellum,
including glial cell activation and changes in Pur­
kinje cells. After TBI, the cerebellum shows pa-
thological changes, including selective cell loss,
altered metabolism, and white matter damage.
Damage to Purkinje cells was thought to be an
indicator of TBI.14,15 In a study conducted in a TBI
group, histopathological findings such as dilation
in capillaries in the granular layer and inflamma­
tory cell infiltration around hemorrhage, Purkinje
cells, some pyknotic nuclei in ganglion cells, de-
generative changes were detected.16 Posttraumatic
cerebellar Purkinje cell damage and loss are
among the important features of cerebellar injury,
including degeneration of glial cells and traumatic
axonal damage.17 Fukuda et al18 investigated an
injury caused by lateral fluid percussion to the
frontoparietal cortex 1 day and 7 days after injury.
They showed that Purkinje cells died in the cere­
bellar vermis at both time points. After trauma,
degeneration and separation of Purkinje cells in
the ganglion cell layer, pyknosis, and apoptotic
Volume 43, Number 4/August 2021 197
Resveratrol in Traumatic Brain Injury
Figure 1 Graphical illustration of GPx values.
Figure 2 Graphical illustration of SOD and MDA values.

6. changes in the nuclei of granular cells were ob-
served. Thrombus and large-diameter vacuolar
structures were seen in the blood vessels of sub­
stantia alba and grisea. It has been observed that
trauma causes degenerative effects on neuron and
glial cells in the cerebellum tissue, as well as
blockages in blood vessels. However, it is thought
that it may affect balance and synaptic function
negatively (Figure 4).
MDA, a lipid peroxidation product, has been
reported to promote the cross-linking of nucleic
acids, proteins, and phospholipids that cause dys-
function of macromolecules, acting as a key mark­
er of lipid peroxidation.19 SOD, an endogenous
antioxidant enzyme, converts harmful superoxide
radicals into hydrogen peroxide. It acts as a super­
oxide radical scavenger to protect the cytoplasm
against damage caused by oxygen-free radicals.20
Resveratrol is a biological antioxidant that can
reduce secondary oxidative stress-induced cell
damage after TBI by increasing serum SOD levels
and decreasing serum MDA levels.21
One study reported that resveratrol administra-
tion after TBI may have a beneficial effect on neu­
rological recovery and antioxidant activity in rats.
When resveratrol administration was compared
with TBI groups, it was shown that resveratrol
significantly increased serum SOD levels in rats
after TBI and decreased serum MDA levels22 (Fig­
ures 1–3). Fu et al23 measured improved biologi­
198 Analytical and Quantitative Cytopathology and Histopathology®
Firidin
Figure 3
Graphical illustration of CAT
values and histopathological
and immunohistochemical
scores.
Figure 4 (A) Control group: normal appearance of Purkinje cells, granular layer, and glial cells (hematoxylin-eosin staining). (B) Trauma
group: degeneration and separation of the ganglion layer, apoptosis in the nuclei of the Purkinje cells. Thrombus and large-diameter
vacuolar structures were seen in the blood vessels in the substantia alba and substantia grisea (hematoxylin-eosin staining). (C) Trauma+
Resveratrol group: a decrease in the degenerative Purkinje cells, an increase in chromatin density in the cells of the granular layer, and an
increase in the acidophilic area in the glomus areas (hematoxylin-eosin staining).

7. cal parameters in spinal cord injury in ischemia-
reperfused rats, measured in spinal cord tissue
homogenates treated with 10 mg/kg resveratrol.
They stated that resveratrol protected the spinal
cord from ischemia damage and significantly
reduced plasma nitrite/nitrate, iNOS mRNA, and
protein expression and p38MAPK phosphoryla-
tion levels in rats, and resveratrol was an impor­
tant antioxidant. Several studies have found that
after treatment of TBI rats with resveratrol, the
expression of Bcl-2 genes associated with inhibi­
tion of apoptosis of neuronal cells is significantly
increased and neuronal apoptosis is significantly
inhibited.24-26
After traumatic brain injury, GFAP, the protein
that serves as the main component of the cyto­
skeleton of astrocytes, plays an important role in
elucidating changes in the blood-brain barrier.
Özevren et al have shown that apoptotic glial cells
are positive in posttraumatic GFAP reaction, sig­
nificant irregularity in astrocytes, and shortened
extensions around the blood vessel.27 It has been
shown that there is a transient correlation between
injury-induced expression of glial fibrillar acidic
protein (GFAP), which is a marker for activated
astrocytes, and the development of neuropathic
pain.28 Cytokine production and GFAP upregula­
tion in astrocytes after injury is thought to be im-
portant in the maintenance of neuropathic pain. It
has been reported that the upregulation of GFAP
was observed 3 days after nerve injury and lasted
until the 21st day.29 After trauma, GFAP expres-
sion was observed in glial footprints around the
blood vessels in the cerebellum and in Purkinje
cells and synapse areas, while upward regulation
of GFAP expression was observed in the molecu-
lar layer and in the synaptic area in the substantia
alba layer in the Resveratrol group. It was thought
that this may be a result of cell protein structuring
due to antioxidative effect (Figure 5A–C).
The importance of ADAMTS proteoglycans has
been predicted to be involved in the neuroin­
flammatory response after CNS injury by pro­
moting infiltration of macrophages into the CNS.
ADAMTS-4 has been shown to play an important
role in macrophage infiltration into the CNS di-
rectly or indirectly after injury by promoting the
leakage of the blood brain/spinal cord barrier.
Regarding the effects of ADAMTS-4 on axonal
regeneration, it was determined that ADAMTS-4
Volume 43, Number 4/August 2021 199
Resveratrol in Traumatic Brain Injury
Figure 5 (A) Control group: moderate GFAP expression in Purkinje cells and weak expression in microglia cells (GFAP immunostaining).
(B) Trauma group: an increase GFAP expression was in the molecular layer of the glial foots, and positive GFAP expression in neuron and
glial cells in the granular and gangliosum layers (GFAP immunostaining). (C) Trauma+Resveratrol group: an increase in GFAP expression
in the neural extensions of the molecular layer and in the synaptic areas (GFAP immunostaining). (D) Control group: positive ADAMTS-4
expression in cells in the granulosa layer, weak ADAMTS-4 expression in glial cells and Purkinje cells. In the trauma group, ADAMTS-4
immunostaining. (E) Trauma group: positive ADAMTS-4 expression in Purkinje cells, glial cells, and extracellular matrix in the glomus
area (ADAMTS-4 immunostaining). (F) Trauma+Resveratrol group: ADAMTS-4 expression Purkinje cells, some of the granular cells, and
in glial cells.

8. reverses proteoglycan-mediated inhibition of neu­
rite growth in vitro and supports functional recov­
ery after TBI.30 In the trauma group, ADAMTS-4
expression was positive in Purkinje cells, glial
cells, and extracellular matrix in the glomus re-
gion, while ADAMTS-4 expression was signifi­
cantly observed in Purkinje cells, some granular
cells, and significantly in glial cells in the Trauma+
Resveratrol group (Figure 5D–F).
Conclusion
In determining the signaling mechanisms involved
in the development of damage in astroglial cells
and other glial cells, Purkinje cells and synaptic
extensions, GFAP and ADAMTS-4 proteins could
provide new therapeutic targets for neurological
diseases. It is thought that it could be helpful to
understand the effect of antioxidative drugs such
as resveratrol.
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