Evaluation of neuroprotective effects of different treatments in
TBI induced Epileptogenesis in rats:
Model standardizing studies confirmed the altered neurobehavioral and
brain complications results in futuristic PTE. Fasudil Hydrochloride
(FH, 10mg/kg i.p.), Flufenamic Acid (FFA, 20mg/kg oral), valproic acid
(VPA, 350mg/kg i.p.), and Valproic Acid + Flufenamic Acid (VFA, 175mg/kg
i.p. + 10mg/kg oral) treatment groups were compared against disease and
normal healthy controls to check the efficacy in TBI progression. TBI
was induced into 6 treatment groups in which 2 were TBI control, EPLT
group, and 4 were selected for treatment studies. One control and one
surgery group i.e. Sham were also considered for comparisons to conclude
the surgery effect. All the groups were analyzed for bodyweight
difference, neurological severity score, and motor coordination on Day0,
Day1, Day3, Day7, and Day14 before proceeding towards molecular
estimation.
Animal Body Weight Difference: The weight severity profile was
observed followed by the head injury. The line graph of weight
difference on Day3, Day7, and Day14 showed a significant difference in
weight difference (grams). All treatment groups i.e. FH, FFA, VPA, and
VFA were found to be significantly effective to maintain the rat weight
compared to disease controls i.e. TBI/EPLT groups on Day7. There was no
significant difference observed between intra-treatment groups on Day7
(Figure-2).
Neurological Severity Scale
Estimation: The NSS observed in
the VFA group was more significant to restoring the neurological score
on Day3, Day7, and Day14 when compared to the TBI and EPLT group. The
other treatment groups showed a significant increase in NSS scores on
Day3, Day7, and Day14 when compared to disease controls (Figure-2).
Rotarod Test for Motor
Co-ordination: The latency to
fall (in seconds) was found to be significant for all treatment groups
when compared to disease controls on Day7 and Day14 (Figure-2).
Seizure Score assessment after Brain Trauma: A sub-convulsive
dose of PTZ(35mg/kg) was injected on day14 and injured rats were
observed for seizure scoring for 2hours. Increased susceptibility for
seizures was confirmed in the EPLT group with a low seizure threshold.
EPLT group showed behavioral arrests, complex partial, dileptic, and
tonic-colonic seizures. But no seizures were observed in any treatment
groups (Table in Figure-2).
Brain Infarction by TTC Staining:The infarction was visible in the
striatum and cortex leaving the hippocampus. Control and Sham group
observed deep red in staining with normal histology with no infarction
in the brain. TBI and EPLT groups were showed hypoperfusion
significantly in the area of infarct (white pale color) when compared to
normal controls. FH, FFA, VFA treatment groups showed a significant
difference to the disease control in red staining i.e. less
hypoperfusion and rich red stain when compared to TBI and EPLT groups.
But VPA significantly shows nearly
red staining of injured brain tissue with very less area of infarct
(Figure-3).
Evan’s Blue
concentration: Evan’s blue (2%,
4mL/kg i.p.) was administered in rats and an abrupt increase of Evan’s
blue was seen in TBI and EPLT groups. But the Evans concentration in FH,
FFA, VPA, and VFA groups was found significantly less compared to injury
controls. VPA alone and in combination with FFA was found very effective
as compared to FH and FFA alone (Figure-3).
Brain Edema difference:Surgically removed rat brains were weighed just after the sacrifice and
after 24 and 48 hours when placed in an incubator at
70oC. The brain
edema in all the treatment groups showed no significant differences when
compared to disease controls after 24 hours but there is a significant
difference after 48 hours i.e. the value of water content in the
treatment groups was found less as compared to the TBI/EPLT groups
(Figure-3).