Para-meter

Controlaspirin

AspirinRanitidine

AspirinOmeprazole

Aspirin-V+R
Gp

0.001242

AspirinVerapamil
0.0483

U.I

0.00071

0.009146

0.010068

pH

0.00070299
1

0.00880385
8

0.03364717
4

0.00013730
4

0.00476424
9

AcidContents

0.00031430
3

0.00099183
5

0.00242060
2

0.00015217
3

0.00061023
3

acetylsalicylic acid (aspirin), Bayer (Leverkusen, Germany)

AQWAL E ZARIAN
Estimation of free acidity

Centrifuge the gastric content at 1000 g for 10 minutes. Note the volume of gastric juice.
Pipette out 1 ml of the supernatant liquid and dilute it to 10 ml with distilled water. Then total
acidity of gastric juice was estimated by titration with 0.01 N sodium hydroxide using
phenolphthalein as an indicator. The result was expressed as the free acid output, which was
expressed as mEq / liter of body weight.

Isolation of stomach and collection of gastric juice

On the twenty-third day (4 h post ulcer induction), the animals were humanely sacrificed
by cervical dislocation. The abdomen was opened and the stomach excised. The
stomach was thereafter opened along greater curvature and gastric content was
drained into a centrifuge tube. Five ml of distilled water was added and the resultant
solution was centrifuged at 3000 rpm for 10 min. The supernatant obtained was
thereafter used for biochemical analyses.

3.7. Determination of gastric secretion parameters

Gastric acid output (volume) was determined in the supernatant (2 ml) by titration with
0.0025 N NaOH using Toepfer's reagent as indicator. The pH of gastric juice was
determined using a pH meter, while the procedures of Sanyal et al. [21] and Corne et al.
[22] were used to determine specific pepsin activity and mucin concentration
respectively.
The

pH gives an idea of the level of acidity and volume of gastric secretions. Low pH value is

a manifestation of decreased hydrogen ion concentration in gastric juice. This has been linked
to pathogenesis of ulcer and gastric damage in experimental animals [31]. Inas et al. [27]

Determination of free acidity total acidity One ml of gastric juice is pipette into
100 ml conical flask, added 2 to 3 drops of topfers reagent and treated with 0.01 N

sodium hydroxide until all traces of red colour disappears and the colour of the
solution turns to yellowish orange. The volume of the alkali added was noted. This
volume corresponds to free acidity. Then 2 to 3 drops of phenolphthalein solution
is added and titration is continued until a definite red tinge reappears. Again the
total volume of alkali added is noted. Acidity is calculated by using the formula

Acidity = volume of NaOH X Normality of NaOHx100/0.1

m.eq/lit/100gms.
3.4 Comparative Study Groups
Group A and Group B: With or without Aspirin to access the extent and
severity of peptic ulcerations induced by Aspirin.
Group B, Group C, Group D and Group E: To observe the degree of gastro
protection offered by Ranitidine, Verapamil and Omeprazole in animals
receiving 2 ml of (75mg/ml) Aspirin.
Group C, Group D, and Group E: To compare the magnitude of gastro
protection offered by Ranitidine, Verapamil and Omeprazole.
Group E and Group F: To compare the magnitude of gastro-protection of
Omeprazole alone with combined effect of Verapamil and Ranitidine.

4.2 Group-B
This group acted as standard to mean U.I of the group 13.83 0.91, the ph was mean
2.92 0.32 and the calculated acid concentration was mean 4.38 2.55. The histopathological
results co-inside with these figures 02 slide show moderate change and four were adjudge to
have severe pathology.

4.3 Group-C
The ulcerprotective effects of Ranitidine was estimated as mean U.I was 11.17 1.18,
the pH mean was 3.59 0.09 and the calculated acid concentration was 0.34 0.07. Percentage
protection was 19.28 % calculated. The histopathology results corresponding the figures and 03
slides were graded as mild and 03 were adjudge in moderate.

4.4 Group-D
Verapamil showed its protective effects against ulcer as mean U.I was 12.48 0.26 ,the
pH was mean 3.22 0.23 and the calculated acid contents concentration was mean 1.23 0.65.
Percentage protection was 9.76 % calculated .The hitopathology reveals that 05 slide were
showing moderate changes and 01 slide was graded as severe pathology.

4.5 Group-E
This group was given Omeprazole, the mean U.I was 9.79 0.35 with pH mean of 4.23
0.28 and estimated acid contents concentration was mean 0.16 0.09. Percentage protection was
29.20 % calculated. The microscopic grading showed 04 slides with no change and 02 slide falls
in the category of mild pathology.

4.6 Group-F
Combined ulceroprotective effects of Ranitidine and Verapamil was perceived as the
mean U.I was 10.89 0.44, the pH mean was 4.04 0.26 and calculated acid contents
concentration was mean of 0.19 0.09. Percentage protection was 21.30 % calculated. The
histopathology reports co-inside with the figures and 04 slide were graded as minimal change
pathology while 02 were adjudge to have moderate pathology.

VERAPAMIL ROLE

Calcium modulators Beginning in 1986, several groups reported an interesting effect of calcium
channel antagonists on experimentally induced gastric ulcers (19, 20). Although these
compounds, including verapamil, nifedipine, diltiazem, and nitrendipine, protect against various
forms of stressinduced gastric lesions, they worsen ethanol-induced gastric mucosal damage,
possibly by reducing gastric mucosal blood flow (19) and possibly through adrenal
glucocorticoid influence (20). In contrast, less selective calcium-modulating compounds, the
calcium chelators EGTA and EDTA, were shown to protect against ethanol-induced gastric
lesions (21). Vagotomy and adrenalectomy abolish the protective effect of EGTA but only
slightly diminished that of EDTA. One possible mechanism to account for the protective effects
of calcium chelators is through inhibition of calcium-dependent proteases or calpains believed to
mediate some forms of gastric mucosal injury (22). One additional mechanism described
recently that may explain the action of calcium chelators to reduce ethanol lesions is inhibition of
conversion of xanthine dehydrogenase to the superoxide-generating enzyme, xanthine oxidase,
shown to occur with EGTA in rat liver (23). A role for oxygen radicals in ischemic mucosal
lesions has been proposed by others (24), although their role in ethanol- or HC1-induced injury is
limited (GARY B. GLAVIN AND SANDOR).
-----------------------------------------------------------------------------------------------------------------

The effect on gastric acid secretion of blocking transmembrane Ca2+ influx into the parietal cells
has been studied in the isolated guinea pig fundic mucosa and in healthy volunteers. Verapamil
inhibited in a dose-related manner histamine-stimulated acid secretion in the guinea pig mucosa,
whereas stimulation with theophylline and dibutyryl cyclic-AMP was unaffected. The effect of
verapamil (Isoptin, 2.0 mg/h) on acid secretion stimulated by increasing doses (50,200, and 500
ng/kg-h) of 15-leucine synthetic human gastrin I was studied in seven healthy volunteers, alone
and in combination with infusion of calcium gluconate (1.0 meq Ca2+/kg-h). Verapamil inhibited
the acid response to the lowest dose of gastrin, resulting in a significant increase of D50 of 15leucine synthetic human gastrin I. This effect was partly reversed by calcium infusion. It is
concluded that one of the mechanisms by which extracellular calcium concentration influences
acid secretion is by transmembrane influx of Ca2+ during stimulation (Calcium and StimulusSecretion Coupling in Gastric Fundic Mucosa by P. Kirkegaarda, J. Christiansena*, B.
Petersena & P. Skov Olsena pages 533-538)-----

Gastroprotective effect of small centaury (Centaurium erythraea L) on

aspirin-induced gastric damage in rats.
Tuluce Y1, Ozkol H, Koyuncu I, Ine H.
Author information

Department of Medical Biology, Faculty of Medicine, Yuzuncu Yil University, Van, Turkey.
yasintuluce@yahoo.com
1

Abstract
The aim of this study was to determine the antiulcer and antioxidant activities of Centaurium erythraea L
(small centaury) in aspirin (ASA) induced acute gastric ulcer model. The gastroprotective effect of the

investigated in rats at a
dose of ASA 200 mg/kg body weight. Twenty-one Sprague-Dawley albino rats
50% aqueous-ethanolic small centaury (SC) extract was

were divided into three groups of seven rats each as follows: (1) control group; (2) acute ASA-treated
group and (3) ASA plus SC group. At the end of the 4-h drug administration, ulcer index, oxidant and
antioxidant levels were measured and compared between the groups. The percentage of lesion area to
total gastric surface area (ulcer index) was significantly reduced (77%) in ASA plus SC group as
compared with acute ASA-treated group. The oral administration of ASA decreased catalase (CAT),
reduced glutathione (GSH), and increased lipid peroxidation (LPO) levels. Although myeloperoxidase
(MPO) activity was increased by ASA, it was found to be lower in the ASA plus SC group. GSH and
Vitamin A levels were determined higher in the ASA plus SC group compared with ASA group. These
results suggest that SC extract protects against ASA-induced damage due to its antioxidizing activity.