Page 89 - AN-4-1

Advanced Neurology Tetrapleura tetraptera protects the hippocampus

Table 1. Schedule of groupings and administration protocol
Groups (n=7) Drugs Dosages (mg/kg) Duration
(alternate days)
Group 1 Control, distilled water 5 mL/kg 11
Group 2 Tetrapleura tetraptera 500 mg/kg 11
Group 3 PTZ 40 mg/kg 11
Group 4 Sodium valproate pre-treatment then PTZ 200 mg/kg then 11
40 mg/kg
Group 5 Low-dose Tetrapleura tetraptera pre-treatment then PTZ 250 mg/kg then 11
40 mg/kg
Group 6 Intermediate-dose Tetrapleura tetraptera pre-treatment then PTZ 500 mg/kg then 11
40 mg/kg
Group 7 High-dose Tetrapleura tetraptera pre-treatment then PTZ 1,000 mg/kg then 11
40 mg/kg
Notes: Pentylenetetrazol was administered intraperitoneally, while Tetrapleura tetraptera fruit extract and sodium valproate were administered orally.
All administrations were carried out every alternate day for 21 days (a total of 11 alternate days).
Abbreviation: PTZ: Pentylenetetrazol.

The excess stain was then washed off with distilled water. 2.12. Statistical analyses
For the immunohistochemical study, antigen retrieval One-way and repeated measures analyses of variance
was performed in a citrate buffer solution (pH 6.0) in a were employed to compare the calculated means and
microwave oven for 5 min, followed by protein blocking percentages. Subsequently, Tukey’s or Bonferroni’s post-hoc
with 3% hydrogen peroxide for 10 min. Sections were tests were performed using GraphPad Prism software
then pre-incubated in 2% normal goat serum for 30 min (version 5.0; United States of America) to determine
and incubated with either a monoclonal mouse anti- statistical significance at p≤0.05. All results are presented as
NSE (22C9, 1:100, Leica Biosystems, United States) or mean ± standard error of the mean or standard deviation.
monoclonal mouse anti-GFAP (NCL-L-GFAP-GA5,
1:100, Leica Biosystems, United States) for 2 h. Afterward, 3. Results
sections were incubated for 1 h with goat anti-mouse
secondary antibody (1:100). The reaction was detected 3.1. Phytochemical analysis of T. tetraptera
using the avidin-biotin complex with diaminobenzidine as Qualitative screening of T. tetraptera fruit extract revealed the
the chromogen. Finally, sections were counterstained with presence of phenols, flavonoids, alkaloids, saponins, tannins,
Harris hematoxylin. terpenoids, steroids, phytate, anthraquinones, oxalates,
All stained sections were dehydrated in increasing cardiac glycosides, and hydrogen cyanide. The quantitative
concentrations of alcohol, cleared in xylene, and analysis of T. tetraptera fruit extract showed that phenols
cover-slipped with dibutylphthalate polystyrene xylene. (7.80 g/100 g) had the highest concentration, followed
Processed slides were examined under a light microscope, by alkaloids (6.20 g/100 g), flavonoids (5.30 g/100 g),
and photomicrographs were captured using a computer- saponins (3.40 g/100 g), tannins (1.20 g/100 g), and phytate
assisted digital microscope camera. (0.40 g/100 g). Oxalates, cardiac glycosides, and hydrogen
cyanide were present in trace amounts.
2.11. Hippocampus cell counts
3.2. Oral median lethal dose
The populations of Nissl substance-stained Cresyl
violet and NSE- and GFAP-immunopositive cells in the Mice that received oral administration of T. tetraptera fruit
hippocampus were quantified manually using ImageJ extract at doses ranging from 1,000 mg/kg to 5,000 mg/kg
®
software (United States of America). Images of the three showed no signs of death or toxicity. Hence, the median
layers of the hippocampal cornu ammonis 3 (CA3) regions lethal dose of T. tetraptera ethanol fruit extract was
were acquired from three brain sections per animal, determined to be greater than 5,000 mg/kg.
with three animals per group. Each image was mapped
randomly using the ImageJ gridlines as a guide. Manually 3.3. Antiseizure activity of T. tetraptera
®
counting was conducted using a tool to select Nissl, NSE, Repeated administration of PTZ at a sub-convulsive dose
or GFAP-stained cells located on the upper and right of 40 mg/kg for 11 alternate days resulted in a steady
borders of each mapped area. increase in seizure scores, leading to generalized clonic-

Volume 4 Issue 1 (2025) 83 doi: 10.36922/an.6862