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Advanced Neurology Pantothenic acid in kainic acid-induced epilepsy

KA (10 mg/kg, i.p.) administration, which was performed was taken at 412 nm for a continued 7-min interval of 1 min.
30 min after the administration of diazepam. The mice, The results were expressed as nM/L/min/gm of tissue.
thereafter, were observed over a period of 4 h for any change in
behavioral parameters, incidence, and latency of convulsions. 2.5.4. GSH estimation
In Groups 4, 5, and 6, KA (10 mg/kg, i.p.) and PA at The GSH level was measured using the Butler method
doses of 30, 60, and 90 mg/kg were given to the groups, with minor modifications. The absorption was
respectively, for 7 days. The mice, thereafter, were observed spectrophotometrically measured at 412 nm (X-Rite 640B
over a period of 4 h for any change in behavioral parameters, spectrophotometer). Different GSH standard concentrations
incidence, and latency of convulsions. were also treated simultaneously to create a standard curve
(1 – 50 μg). Results were reported as GSH nmol/mg protein .
[19]
2.5. Behavioural tests
2.5.5. Catalase estimation
2.5.1. Single-trial passive avoidance test
Catalase activity was measured using a previously described
Memory retention deficit was evaluated by step through method , wherein the breakdown of H O was measured.
[21]
2
2
passive avoidance apparatus. The apparatus consists of equal The processed sample homogenate absorbance was taken
sized light and dark compartments (30 × 20 × 30 cm). A 40-W at 240 nm. The results were expressed as micromoles of
lamp was fixed 30 cm above its floor in the center of the light H O decomposed/min/mg/of protein.
compartment. The floor consists of metal grid connected to 2 2
a shock scrambler. The two compartments were separated 2.5.6. SOD enzyme activity
by a trap door that could be raised to 10 cm. Twenty-four h The SOD activity was measured with the enzyme kit from
after the administration of KA, mice were placed in the light RANDOX Ransod. This procedure uses xanthine and
compartment and the time lapse before each animal entered
the dark compartment and had all four paws inside it was xanthine oxidase-produced radicals of superoxides that
measured in seconds and termed as “initial latency” (IL). react with the red formazan dye using 2-(4-iodophenyl)-3-
Immediately, after the mice entered the dark chamber with (4-nitrophene)-5-phenyltetrazolium-chloride. The degree
[22]
all the four paws inside the dark chamber, the trap door was of inhibition of the SOD activity was measured .
closed and an electric foot shock (50 V a.c.) was delivered for 2.5.7. Nitrites content estimation
3 s. Five s later, the mice was removed from the dark chamber
[23]
and returned to its home cage. Twenty-four h after the IL, It was measured by previously described method .
the latency time was measured again in the same way as in 0.2 mL of supernatant from the brain homogenate was
the acquisition trial and was termed as the retention latency mixed with freshly prepared Griess reagent solution and
(RL). However, during the retention trial, no foot shock was the absorbance was measured at 546 nm. The results were
delivered, and the latency time was recorded to a maximum calculated as nM/mg of protein.
of 600 s. To improve the reliability and validity of the foot 2.5.8. Determination of MAO activity
shock avoidance test, the grid as well as the mice paw was
moistened with water before delivering the foot shock as this MAO activity was assessed by spectrophotometry. The assay
is known to reduce the wide inter animal variability in paw mixture contains 4 mm of serotonin as a specific substrate
skin resistance of the mice . for MAO-A, 250 μL solution of brain homogenate, and
[20]
100 mm of sodium phosphate buffer (pH 7.4) up to a final
2.5.2. Evaluation volume of 1 mL. The action was allowed to proceed at 37°C
The animals were sacrificed by cervical dislocation. The for 20 min and stopped by adding 1 M HCl (200 μL). The
whole brain was removed from the skull immediately reaction product was extracted with 5 mL of butyl acetate.
after dislocation. For preparation of the homogenate, the Blank samples were prepared by adding 1 M HCl (200 μl)
fresh whole brain was weighed and transferred to a glass before the reaction and worked subsequently in the same
[24]
homogenizer and 10% (w/v) tissue homogenates were manner .
prepared in 0.1 M phosphate buffer (pH 7.4, stored at
2 – 8°C). The homogenate was centrifuged at 3000 rpm 2.5.9. Histopathology study of the rat brain
for 10 min and the resultant cloudy supernatant liquid was The cortex and hippocampus of control and experimental
used for various neurochemical estimations. rats were fixed in 4% formalin and embedded in paraffin.
Next, they were sliced into 5 μm sections using a section
2.5.3. Choline esterase enzyme determination cutter (Leica, Germany). The sections were stained
The enzyme level was measured by the method described by with hematoxylin and eosin and examined under a light
Ellman with slight modifications. The sample absorbance microscope.
[13]
Volume 1 Issue 2 (2022) 3 https://doi.org/10.36922/an.v1i2.40