Page 109 - ITPS-7-1

INNOSC Theranostics and
Pharmacological Sciences Anti-ulcer activity of Rheum spiciforme

Group I was the control group that was given a vehicle 0.01 N NaOH was performed until a yellow coloration
once per day for 7 days. Group II was the reference group appeared. The volume of 0.01N NaOH used was noted. The
administered with 20 mg/kg omeprazole once per day for total and free acidity was calculated using the following
7 days. Groups III, IV, and V were treated with butanol, formula .
[17]
aqueous, and dichloromethane fractions of plant extract V × N 1 00 mEq / L
×
at 250 mg/kg once per day for7 days. Then, gastric ulcer Acidity = NaOH (III)
was induced in adherence to protocols briefly mentioned 0.1
in section 2.5 . where V NaOH = volume of NaOH solution used, and
[15]
2.7. Macroscopic evaluation of ulcer N= normality.

2.7.1. Estimation of ulcer severity 2.7.6. Determination of gastric juice volume
Stomach of each rat was cut open with an incision on the Gastric juice was centrifuged at 500 rpm for 5 min, and
greater curvature. After the gastric contents were removed, then, the supernatant was separated and its volume was
[20]
gastric lesion on the stomach was examined with the naked measured using a graduated cylinder .
eyes. The length of the ulcer was measured using Vernier 2.8. Liquid chromatography-mass spectrometry
Calliper, and the ulcer severity was estimated by means of (LC-MS) analysis of bioactive fractions
scoring .
[16]
In the present study, active constituents in most active
2.7.2. Estimation of ulcer index fractions (butanol fractions) were identified by means
The ulcer index is defined as mean ulcer severity score of LC-MS using methanol (100%) as the mobile phase.
for each animal. The ulcer index was calculated using the A thermo electron (LTQ-Orbitrap XL mass spectrometer)
[17]
following formula : equipped with a nanoelectrospray ion source (Thermo
Fisher Scientific, Bremen, Germany) was utilized in positive
UI = UN+US+UP×10−1 (I) and negative ionization modes as per requirements.
where UI = ulcer index, UN = average number of The separated target compounds were detected in direct
ulcer per animal, US= average severity score, and UP = injection mode. The collision-induced dissociation (CID)
percentage of animal with ulcer was based on the parent ion fragmentation efficiency. The

2.7.3. Percentage inhibition of ulcer capillary temperature was set at 280°C and the sample flow
at 8 µL/min .
[21]
Percentage inhibition of ulcer is also known as percentage
protection. Percentage inhibition of ulceration was 2.9. Statistical analysis
calculated using the following formula : Data were analyzed using GraphPad Prism, and descriptive
[18]
(UI − UI ) 100× statistics were used to describe the results. The results are
=
Inhibition of ulceration Ctrl Test (II) expressed as mean ± standard error of mean. One-way
UI Ctrl analysis of variance (ANOVA) followed by Dunnett and
where UI Ctrl = Ulcer index of control group, and Bonferroni’s post hoc test was carried out. A P-value <0.05
UI = ulcer index of test group. was considered statistically significant.
Test
2.7.4. Evaluation of pH 3. Results
Before pH measurement, 1 mL gastric juice was diluted 3.1. Gastro-protective effect of R. spiciforme on
with 1 mL of distilled water, and then, pH meter was used gastric mucosal injury in ethanol-induced ulcer rat
to measure the pH of the solution . model
[19]
The extract at a dose of 125 mg/kg resulted in a non-
2.7.5. Determination of free and total acidity
significant decrease (6.50 ± 0.95) in number of lesions in
One milliliter of gastric juice solution, diluted in a 1:1 comparison with the control group (7.83 ± 0.47), whereas
ratio with distilled water, was poured into a conical flask. a remarkable reduction in the number of lesions in mucosa
A few drops of a phenolphthalein indicator were added to was observed when the 250 and 500 mg/kg doses were used
the solution. Titration was performed with 0.01 N NaOH (2.50 ± 0.4 and 1.66 ± 0.4, respectively). The 125, 250, and
until a sustained pink coloration appeared. The volume of 500 mg/kg doses of the plant extract considerably reduced
0.01 N NaOH used was noted. Topfer reagent was used to the ulcer severity score (22.67 ± 7.3, 7.16 ± 1.70, and
determine the free acidity. Titration of gastric juice with 6.83 ± 1.77) compared to the control group (32.33 ± 2.81).

Volume 7 Issue 1 (2024) 3 https://doi.org/10.36922/itps.1343