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Effect of Lemongrass and Green tea on blood pressure and heart rate


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Objective: Lemon grass and Green tea are the herbal products, widely used, next to that of water. Because of their common use, it is so much necessary to find their effects on all the body functions. In the current study its effects of blood pressure and heart rate of human male has been evaluated. Study Design: Observational study. Place of Study: This study was conducted at Pharmacy Department, University of Malakand. Materials and Methods: Seventy two male volunteers for each tea had been selected and the blood pressure before and after giving one cup of each tea to each individual was evaluated by using sphygmomanometer and stethoscope. Results: A minor increase in blood pressure was noted in the volunteers taken green tea. On other hand a moderate decrease in the systolic blood pressure and mild increase in the diastolic blood pressure were noted in the case of lemon grass. Also, great decreases in the heart rate of individuals taken lemon grass, and a moderate increase in the heart rate of individuals taken green tea were observed. Conclusion: From the current study it can be concluded mat the Male heart patients are on the high risk to the use of either type of tea, so they have to take care while using either type of tea in excess quantity.
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Drug and Chemical Toxicology
ISSN: 0148-0545 (Print) 1525-6014 (Online) Journal homepage:
Protective effect of the solvent extracts of
Portulacca oleracea against acidified ethanol
induced gastric ulcer in rabbits
Muhammad Shah Zeb Jan, Waqar Ahmad, Abdullah, Atif Kamil, Mir Azam
Khan, Maqsood Ur Rehman, Irfan ullah & Muhammad Saeed Jan
To cite this article: Muhammad Shah Zeb Jan, Waqar Ahmad, Abdullah, Atif Kamil, Mir Azam
Khan, Maqsood Ur Rehman, Irfan ullah & Muhammad Saeed Jan (2019): Protective effect of the
solvent extracts of Portulacca�oleracea against acidified ethanol induced gastric ulcer in rabbits,
Drug and Chemical Toxicology, DOI: 10.1080/01480545.2019.1691584
To link to this article:
Published online: 19 Nov 2019.
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Protective effect of the solvent extracts of Portulacca oleracea against acidified
ethanol induced gastric ulcer in rabbits
Muhammad Shah Zeb Jan
, Waqar Ahmad
, Abdullah
, Atif Kamil
, Mir Azam Khan
, Maqsood Ur Rehman
Irfan ullah
and Muhammad Saeed Jan
Department of Pharmacy, University of Malakand, Chakdara, Pakistan;
Department of Biotechnology, Abdul Wali Khan University
Mardan, Pakistan
Portulacca oleracea L. has been used for treatment of different ailments. The aim of this study was to
investigate the effectiveness and possible mechanism of action involved in the anti gastric ulcerogenic
effect of Portulacca oleracea. Methanolic extract & subsequent fractions (100, 200 and 400 mg/kg) of
Portulacca oleracea (P. oleracea) were administered orally to experimental rabbits one hour before oral
administration of HCl/ethanol (40:60). Anti gastric ulcerogenic potential of P. oleracea was evaluated by
assessment of gastric pH, pepsin, free acidity, ulcer index, mucus content and total acidity. For the
investigation of possible mechanism of action malondialdehyde (MDA), histamine, and H þKþATPase
content were determined in the stomach homogenate. Histopathological study of stomach tissue was
carried out by H&E dye. Ethyl acetate fraction (EAF) of P. oleracea was the most potent fraction among
all fractions that exhibited efficient protection against acidified ethanol mediated gastric-ulcer. The
ethyl acetate fraction (EAF) significantly increased the pH of gastric juice, while pepsin and histamine
was observed to decrease significantly in comparison to acidified ethanol group (p0.001). The
EAF showed moderately H þKþATPase inhibitory activity. Moreover, it was also observed that EAF
decreased the malondialdehyde (MDA) level in the stomach tissue homogenate showing antioxidant
effect. Histopathological studies showed that among the tested fractions, EAF significantly prevented
acidified ethanol induced gastric mucosal damage. These results showed that mechanism of anti gas-
tric ulcerogenic potential of P. oleracea could be associated with the reduction in histamine level,
HþKþATPase inhibition and reduced MDA level.
Received 23 May 2019
Revised 31 October 2019
Accepted 2 November 2019
Portulacca oleracea; gastric
ulcer; antioxidant;
histamine; pepsin and
Hþ-K þATPase
Ulcer is an injury or sore in the mucous membrane or outer
surface skin of the body. Ulcer in the lining of the stomach
or duodenum is a disease of digestive system that affect
many people around the world (S
anchez-Mendoza et al.
2011). It has been documented that fourteen million people
throughout the world are suffering from gastric ulcer with a
mortality rate of four million. Gastric ulcer occur as a result of
imbalance between aggressive (alcohol, pepsin and acid
secretion, poor diet, oxidative stress, NSAIDs and Helicobacter
pylori) and protective factors (mucosal blood flow, mucus
secretion, bicarbonate secretion and increased levels of anti-
oxidants etc.) in the stomach (Zakaria et al. 2016b). Gastric
mucosa is damaged when aggressive factors overcome
mucosal defensive mechanisms (Laine et al. 2008).
The ethanol-induced ulcer model is widely used for inves-
tigating the testing agents or studying the efficacy of poten-
tial drugs (Strasser et al. 2014). It is well known that ethanol
increase histamine secretion in the stomach which in turn
release hydrochloric acid from the parietal cells inside
mucosa of gastric tissue via proton pump (Kumar and Kumar
2009). A membrane-bound enzyme (H
ATPase), located in
the canalicular membrane of parietal cells, catalyzes an elec-
troneutral exchange of H
versus K
into the gastric lumen
with the expenditure of ATP (Reyes-Chilpa et al. 2006). It is
documented that gastric ulcer induced by ethanol causes
reactive oxygen species (ROS) production because ethanol
rapidly reaches the gastric mucosa and solubilize the protect-
ive mucous and finally releases free radicals and superoxide
anion. These free radicals react with lipid to form malondial-
dehyde (MDA), a marker of lipid peroxidation (Zakaria et al.
2011). The treatment of gastric ulcer has become a challenge
in the world. Although there are some approaches used for
treatment of gastric ulceration like acid neutralization,
improving antioxidant level in the stomach but the most
important approach is inhibition of gastric acid secretion
inside the stomach (Halim et al. 2017).
The current drugs available in the market for the treat-
ment of gastric ulcers are histamine inhibitors (H
such as cimetidine, ranitidine etc.), proton pump inhibitors
(esomeprazole, omeprazole etc.) and mucosal protective
agents like sucralfate and bismuth compounds (Shu et al.
2013). But these drugs have many undesirable effects on the
human body including gynecomastia, arrhythmia and
CONTACT Waqar Ahmad Department of Pharmacy, University of Malakand, Chakdara, Khyber Pakhtunkhwa, Pakistan
ß2019 Informa UK Limited, trading as Taylor & Francis Group
hematopoietic changes (Zakaria et al. 2016a). Therefore medi-
cinal plants are important alternative for the development of
new drugs to control gastric ulcer (Al-Wajeeh et al. 2016).
P. oleracea is an annual herb from family Portulacaceae. It
is widely distributed throughout the world (Okafor et al.
2014). This plant has been folklorically used for the treatment
of many diseases such as diarrhea, bacillary dysentery, diur-
etic, hemorrhoids, emollient, anthelmintic, antiscorbutic
(Guichard 2013) and as gastric sedative (Dalziel 1937). P. oler-
acea has been reported to have various pharmacological
activities like analgesic and anti-inflammatory (Chan et al.
2000), wound healing (Rashed et al. 2003), hepatoprotective
(Anusha et al. 2011), antitumor (Zhao et al. 2013), antioxidant,
hypoglycemic and pancreatic protective activity (Ramadan
et al. 2017). Different phytochemicals have been detected in
P. oleracea include flavonoids, fatty acids, alkaloids, polysac-
charides, terpenoids, vitamins, proteins, sterols and minerals
(Zhou et al. 2015). Many Flavonoids have been reported to
possess gastroprotective activity (Mota et al. 2009). Many
plants having saponins have been reported to possess gas-
troprotective activity (Ugwah et al. 2013). According to litera-
ture, ethanolic extract of P. oleracea has been reported to
have gastroprotective activity against HCl/ethanol induced
gastric ulcer in animal models (Gholamreza et al. 2004).
Rabbits are important animal models used for studying
gastroprotective activity (Okwuosa et al. 2011). Mammalian
stomachs possess histological similarities among species. On
the basis of the gastric wall, rabbits have a predominant
population of chief cells like humans stomach. Thus, rabbit
shares more structural similarities with human. The thickness
of human mucosa is 1 mm which is larger than rats and mice
mucosa. So rabbits are more suitable animal models for gas-
tric ulcer study (Maeng et al. 2013). Therefore on the basis of
folk use, saponins, flavonoids contents and reported in vivo
study, the current study was conducted to elucidate the anti
gastric ulcer potential of all fractions and the possible mech-
anism of P. oleracea.
Material and methods
Chemicals and drugs
Ranitidine (Cirin Pharmaceuticals Hattar Pakistan), Absolute
ethanol (Fisher Scientific, Loughborough, UK), HCl (Lab-Scan,
Analytical Sciences, Pathumwan, Bangkok), Bovine albumin,
Adenosine triphosphate (ATP), Thiobarbituric acid, Histamine
(Sigma-Aldrich, St. Louis, USA), O-phthal-aldehyde (JebChem,
Bangkok,Thailand), Trichloroacetic acid, Methyl orange, Folin
Ciocalteau reagent, Sodium carbonate, Magnesium chloride,
Potassium chloride, Sodium hydroxide and Ammonium
molybdate (Merck, Darmstadt, Germany), n-Butanol and
Phenolphthalein (BDH), Perchloric acid (Scharlu, Sentmenat,
Spain). Chloroform, n-hexane, ethyl acetate and methanol
were of commercial grade.
Plant material
Mature plant of P. oleracea was collected from Charsadda,
Khyber Pakhtunkhwa, Pakistan. Identification of this was
made by botanist (Dr. Nasrullah Khan) at University of
Malakand (Department of Botany) and deposited in herbar-
ium with voucher No. H.UOM.BG.174.
Extraction and fractionation
The plant was air dried in shady area for 60 days at room
temperature followed by crushing via cutter mill into powder
(5 kg) and subsequent soaking in 80% methanol for 3 weeks.
The plant extract was filtered with the help of muslin cloth
and evaporated by using the rotary evaporator to obtain the
methanolic extract of P. oleracea (MEE, 360 g). Methanolic
extract (300 g) was further fractionated with solvents like
chloroform, n-hexane, ethyl acetate and water (Zeb et al.
2014). The chloroform (CHF), ethyl acetate (EAF), n-hexane
(NHF), and aqueous fraction (AQF) obtained were, 18, 27, 43,
and 98 g respectively.
Adult rabbits (1 to 1.5 kg) of either sex were bought from
local market. Animals were kept in the animal house at
University of Malakand, with twelve hours dark and light
cycle having free access to water and standard diet. Before
the start of experiment, animals were fasted overnight while
water was withdrawn 2 h earlier. The experimental proce-
dures were approved by the Ethical Committee of the
Department of Pharmacy according to Animal Bye Laws 2008
of the University of Malakand (NO.Pharm/ECC/PO-142/12/17).
Acute oral toxicity
Rabbits were randomly allocated into 18 groups, each contain-
ing 6 animals. Crude extract was dissolved in distilled water
and was administered orally at various doses ranging from 5
to 2000mg/kg. After receiving the doses, the rabbits were
observed for next 72 h for mortality/minor abnormal behavior
as well as any other allergic symptoms (OECD 2000).
Induction of gastric ulcer using HCl/ethanol
Rabbits were grouped into 18 (n¼6). Group 1 was adminis-
tered distilled water (10 ml/kg p.o). Groups 24 were given
with 100, 200 and 400 mg/kg doses of liquid MEE orally.
Groups 57 were given with 100, 200 and 400 mg/kg doses
of liquid NHF orally.
Groups 810 were given with 100, 200 and 400 mg/kg doses
of liquid CHF orally.
Groups 1113 were given 100, 200 and 400 mg/kg doses of
liquid EAF orally.
Groups 1416 were given 100, 200 and 400 mg/kg doses of
liquid AQF orally.
Group 17 (Ranitidine, 50 mg/kg orally) and XVIII
(Omeprazole 20 mg/kg orally) were used as positive control
(for H
ATPase only). After 60 min each animal was given
1 ml/250 g of weight HCl/ethanol solution (150 mM HCl in
2 M. S. Z. JAN ET AL.
60% ethanol) orally. After two hours animals were euthanized
using diethyl ether and were sacrificed (Ateufack et al. 2015).
Measurement of ulcer index
The stomachs of rabbits were pierced from the site of greater
curvature and gastric contents were removed. The stomachs
were then washed with distilled water to examine ulcers in
the glandular portion of the stomach. The number of ulcers
per stomach was calculated with the aid of magnifying lens.
The scoring was given as 0 for normal stomach, 0.5 for red
coloration, 1 for spot ulcer, 1.5 for hemorrhagic streaks, 2 for
ulcer 35 mm and 3 for ulcer >5 mm. The ulcer index for
each animal was expressed as mean ulcer score (Chetan
et al. 2013).
Estimation of free acidity, total acidity and pH of
gastric juice
The gastric content obtained from dissected stomach was
centrifuged at 2500 rpm for 10 min to remove solid material.
The pH of the supernatant of gastric juice was measured
with help of pH meter. For the measurement of free acidity
of gastric juice, 23 drops of methyl orange reagent was
added to the gastric juice and titrated with 0.01 N NaOH till
the appearance of pale color. The volume of added NaOH
was noted. Then, 23 drops of phenolphthalein were added
to the same solution for measurement of total acidity. The
titration was carried out until pink color appeared. The total
volume of NaOH added to the solution was noted. Acidity
was measured with the help of the following formula (Katary
and Salahuddin 2017).
Acidity ¼Volume of NaOH Normality of NaOH
0:1100 meq=L
Estimation of gastric mucus production
The gastric mucosal layer of each rabbit was smoothly
scraped off with the aid of glass slide and the mucus
obtained was weighed via electronic balance and compared
with positive control (Sidahmed et al. 2013).
Determination of pepsin
Gastric juice (1 ml) was taken in test tube with 1 ml albumin
solution (0.5% BSA w/v in 0.06 N hydrochloric acid) and was
incubated at 37 C for 10 min. For the termination of the
reaction, 10% trichloroacetic acid (TCA) 2 ml was added and
centrifuged the sample at 1500 rpm for 20 min. Sodium car-
bonate 0.55 M (2.5 ml) and 1 ml of Folins reagent (FCR) were
added to the tubes containing supernatant and incubated at
room temperature for 30 min. The absorbance of each sam-
ple was determined via spectrophotometer at 660 nm. The
activity of pepsin was represented as mg of tyrosine/ml
(Moram et al. 2016).
Histopathological study
Stomach from each rabbit were removed and washed with
normal saline solution. The excised stomach of each group
was kept in 10% neutral buffered formalin. The specimen of
gastric tissue was placed in paraffin wax. The specimen was
then stained with hematoxylin and eosin dye and examin-
ation was carried out with the help of light microscope for
the assessment of histopathological changes (Laloo
et al. 2014).
Mechanistic activity of P. oleracea
Estimation of malondialdehyde (MDA) level
One gram of gastric tissue was homogenized in phosphate
buffer and incubated at 37 C. Homogenized gastric tissue
(1 ml) was mixed with 10% TCA (1 ml) and then centrifuged
for a period of 20 min at 2000 rpm. The supernatant was
mixed with 2 ml of 0.67% TBA and heated for 30 min at
100 C in a water-bath. After cooling, the suspension was
centrifuged at 2000 rpm for 10 min. The absorbance of pink
colored supernatant was read via spectrophotometer at
532 nm. The result of MDA content was expressed as nano
moles per gram of wet tissue (Ping et al. 2005).
Estimation of histamine content
One gram of stomach tissue was homogenized with the aid
of Teflon homogenizer in 0.4 N perchloric acid followed by
centrifugation at 3000 rpm for 10 min. The n-butanol was
used for extraction of histamine from supernatant and then
n-butanol was mixed with 2 ml of 0.1 N HCl and 15 ml of
n-heptane. After this, centrifugation at 2000 rpm for 10 min was
carried out. The heptane layer was removed and discarded
while the HCl layer was mixed with o-phthal-aldehyde (OPA) for
the formation of fluorescent product. Spectrofluorimeter
(450 nm emission & 360 nm activation) was used for the meas-
urement of histamine. Result was expressed as micro gram of
histamine released/g tissue (Laloo et al. 2013).
Determination of H
Proton potassium ATPase was obtained by scrapings rabbit
gastric mucosa. One gram of mucosa was homogenized with
the help of homogenizer in 100 ml of Tris-HCl buffer and cen-
trifuged at 5000 rpm for 10 min. The resulting supernatant
was recentrifuged at 5000 rpm for 20 min. The protein con-
centration in the supernatant was determined using bovine
serum albumin as standard (Lowry et al. 1951). One ml reac-
tion mixture containing enzyme was mixed with 20 mM Tris-
HCl (0.2 ml) having pH 7.4, 2 mM MgCl
and 2 mM of KCl
(0.2 ml). Reaction was started by adding of 0.2 ml of 2 mM
ATP (adenosine triphosphate) and incubated at 37 C for
30 min. The reaction was terminated by adding one ml of
TCA and 0.5 ml ammonium molybdate. The sample was cen-
trifuged for 10 min at 2000 rpm. The absorbance was meas-
ured with spectrophotometer at 640 nm to determine the
release of inorganic phosphorus from ATP. The enzyme
activity was expressed as nM of phosphate liberated/min/mg
protein (Selvamathy et al. 2010).
Statistical analysis
The results of all assays were expressed as mean ± standard
error mean (SEM). One Way Analysis of Variance (ANOVA)
was used for analysis of data followed by post hoc Dunnetts
test for multiple comparisons of data. All the values were
considered significant when p<0.05.
Acute oral toxicity
In the acute toxicity study, no mortalities were recorded in
animals treated with a single dose of 2000 mg/kg body
weight. There were no clinical signs in the eyes and mucus
membrane, skin and fur, respiratory rate, autonomic effects,
circulatory signs, and central nervous system at dose
2000 mg/kg body weight. Therefore, the approximate lethal
dose in the experimental rabbit was higher than 2000 mg/kg.
According to organization for economic cooperation and
development (OECD) guidelines for acute oral toxicity, an
LD50 dose of >3002000 is categorized as category 4 and
hence the plant extract is found to be safe.
Macroscopic evaluation
The anti gastric ulcer activity of P. oleracea in HCl/ethanol-
induced gastric lesion model is shown in Figure 1. Ethanol/
HCL solution cause damaged to the gastric mucosa of rab-
bits. Different fractions of P. oleracea significantly reduced
the formation of the gastric lesion. Based on our result, EAF
was the most potent fraction and it reduced the gastric
mucosal ulcer that was comparable with raniti-
dine (standard).
Effect on gastric ulcer index
The effects of the crude methanolic extract and subsequent
fractions of P. oleracea in the acidified ethanol-induced gas-
tric ulcer models are shown in Figure 2. The results show
that the EAF (3.3 ± 0.54) significantly reduced the Ulcer index
at highest dose (400 mg/Kg), when compared to the negative
control group (30 ± 0.44). These results suggest that EAF
demonstrated better healing of the lesions induced by acidi-
fied ethanol than other groups.
Effects of P. oleracea on pH, free and total acidity of
gastric juice
The effects of crude methanolic extract and subsequent frac-
tions of P. oleracea on acidity of gastric juice of rabbit after
administration of acidified ethanol are shown in Table 1.
Among the fractions, the EAF significantly increased the pH
and decreased the free and total acidity of gastric juice of
rabbit stomach as compared to negative control group.
Effects of different fractions of P. oleracea on
mucus production
Rabbits pretreated with crude methanolic extract and subse-
quent fractions of P. oleracea increased the mucus secretion
as shown in Table 2. The negative control decreased the
mucus secretion as compared to standard drug. The EAF was
the most potent fraction and significantly increased the
mucus secretion closely related to positive control group.
Effects of P. oleracea on peptic activity
Negative control showed significant increase in the pepsin
content. The crude extract and subsequent fractions of
P. oleracea decreased the pepsin secretion after administra-
tion of acidified ethanol as shown in Table 3. The result
showed that the most potent fraction (EAF) significantly
inhibited the pepsin release inside the rabbit stomach closely
related to positive control group.
Effects of different fractions of P. oleracea on MDA
The negative ulcerated control showed an increased level of
MDA in the gastric tissue. Pretreatment with crude extract
and subsequent fractions of P. oleracea decreased the MDA
level as shown in Table 4. The results showed that the EAF
significantly decreased in the MDA level closely related to
standard drug (ranitidine).
Effects of different fractions of P. oleracea on
histamine level
The histamine level of rabbit gastric tissue of ulcer negative
control group was significantly increased as compared to
other groups. Pretreatment with crude extract and subse-
quent fractions of P. oleracea decreased the level of hista-
mine in gastric tissue as shown in Figure 3. The EAF
significantly decreased the level of histamine in gastric tissue
than other groups, closely related to positive control group.
Effects of different fractions of P. oleracea on H
ATPase enzyme
Rabbits treated with a standard drug (omeprazole) showing a
significant inhibitory effect on H
ATPase enzyme by
reducing the release of phosphate ions (Pi) as compared to
negative control group. The EAF of P. oleracea at high dose
(400 mg/kg) showed moderate inhibitory effect on H
ATPase enzyme as compared to standard drug. MEE and CHF
showed very moderately inhibitory effect on H
enzyme as shown in Figure 4.
4 M. S. Z. JAN ET AL.
Histological evaluation
Histological observation using H&E staining confirm the abil-
ity of crude and subsequent fractions of P. oleracea to pre-
vent gastric damage of the gastric mucosa induced by
acidified ethanol (Figure 5). The negative control group
showed highly extensive gastric mucosal damage (black
arrow), edema (white arrow) and leucocytes infiltration (blue
arrow) as shown in (Figure 5(B)). Among different fractions
the EAF (Figure 5(CE)) and Ranitidine (Figure 5(A)) confined
gastric tissue from the effect ethanol/HCL having very mild
mucosal disruption, no leucocytes infiltration and edema in
comparison to negative control group.
Figure 1. Effect of P. oleracea on gross appearance of the gastric mucosa in rabbits. (A1) Positive control group (ranitidine 50 mg/kg) protected gastric mucosa. (B1)
Gross appearance of group pretreated with distilled water (ulcer control). Extensive damage of the gastric mucosa was observed. (C1, D1 and E1) Rabbits pretreated
with EAF (100, 200 and 400 mg/kg) þAcidified ethanol reduced gastric lesion comparable to positive control.
Acidfied ethanol
100 mg/ kg
200 mg/kg
400 mg/kg
50 mg/Kg
Ulcer Index
Figure 2. Effect of P. oleracea on ulcer index of rabbits. Values are represented as
mean and n¼6, p<0.001, p<0.01 and p<0.05 as compare to control.
Table 1. Effect of P. oleracea on gastric juice parameters.
Group Dose mg/kg pH
Free acidity
Total acidity
HCl/Ethanol 1.36 ± 0.10 121.33 ± 0.88 133 ± 1.26
Ranitidine 50 4.96 ± 0.18 15 ± 0.54 25 ± 0.54
100 3.06 ± 0.03 71.0 ± 1.14 84.4 ± 0.92
Methanolic 200 3.18 ± 0.16 66.0 ± 2.35 79.0 ± 0.63
400 3.86 ± 0.12 52.2 ± 1.16 64.2 ± 2.15
100 1.52 ± 0.14 85.4 ± 1.21 106 ± 1.58
n-hexane 200 1.88 ± 0.16 80.2 ± 1.02 97.0 ± 1.82
400 2.07 ± 0.01 74.8 ± 0.97 91.6 ± 1.29
100 2.40 ± 0.09 78.0 ± 0.95 94.2 ± 1.39
Chloroform 200 2.71± 0.03 69.2 ± 0.97 85.2 ± 1.24
400 2.98 ± 0.06 64.4 ± 1.66 79.6 ± 1.33
100 4.38 ± 0.07 42.0 ± 1.26 59.0 ± 1.87
Ethyl acetate 200 4.50 ± 0.05 39.6 ± 1.75 47.2 ± 1.11
400 4.64 ± 0.08 22.4 ± 0.98 30.0 ± 1.22
100 4.11 ± 0.11 55.4 ± 0.51 67.4 ± 1.43
Aqueous 200 4.29 ± 0.05 46.2 ± 0.80 57 ± 1.52
400 4.56 ± 0.18 36.0 ± 1.30 49 ± 1.14
Values are represented as mean and n¼6, p<0.001, p<0.01 and
p<0.05. as compared to standard drug ranitidine.
Table 2. Effect of P. oleracea on mucus content.
Group Dose mg/kg Mucus content (gram)
HCl/Ethanol 1.45 ± 0.195
Ranitidine 50 4.60 ± 0.103
100 3.39 ± 0.092
Methanol 200 3.52 ± 0.135
400 3.79 ± 0.104
100 2.07 ± 0.217
n-Hexane 200 1.94 ± 0.145
400 1.74 ± 0.200
100 2.13 ± 0.164
Chloroform 200 2.77 ± 0.207
400 3.32 ± 0.155
100 3.92 ± 0.098
Ethyl acetate 200 4.29 ± 0.143
400 4.47 ± 0.117
100 3.56 ± 0.125
Aqueous 200 3.89 ± 0.200
400 4.21 ± 0.088
Values are represented as mean and n¼6, p<0.001, p<0.01 and
p<0.05 as compared to standard drug ranitidine.
Table 3. Effect of P. oleracea on peptic activity.
Group Dose mg/kg
Pepsin activity
(mg Tyrosine released/ml)
HCl/Ethanol 28.32 ± 0.30
Ranitidine 50 5.30 ± 0.95
100 16.84 ± 0.24
Methanol 200 14.92 ± 0.22
400 13.20 ± 0.46
100 24.82 ± 0.93
n-Hexane 200 22.23 ± 0.57
400 22.34 ± 0.26
100 19.83 ± 0.73
Chloroform 200 18.75 ± 0.67
400 17.12 ± 0.39
100 10.70 ± 0.28
Ethyl acetate 200 9.80 ± 0.11
400 8.38 ± 0.40
100 15.63 ± 0.05
Aqueous 200 13.84 ± 0.64
400 12.16 ± 0.36
Values are represented as mean and n¼6, p<0.001, p<0.01 and
p<0.05 as compared to standard drug ranitidine.
Table 4. Effect of P. oleracea on MDA.
Group Dose mg/kg MDA nmol/g tissue
HCL/Ethanol 78.2 ± 0.30
Ranitidine 50 12.08 ± 0.49
100 46.76 ± 0.62
Methanol 200 34.28 ± 0.55
400 28.17 ± 0.47
100 76.96 ± 0.70
n-Hexane 200 70.48 ± 0.62
400 64.13 ± 0.73
100 57.06 ± 0.86
Chloroform 200 48.11 ± 0.87
400 38.80 ± 0.64
100 38.61 ± 0.49
Ethyl acetate 200 28.32 ± 0.30
400 17.16 ± 0.27
100 40.62 ± 0.28
Aqueous 200 30.15 ± 0.35
400 20.06 ± 0.42
Values are represented as mean and n¼6, p<0.001, p<0.01 and
p<0.05 as compared to standard drug ranitidine.
11 400 mg/kg
200 mg/kg
100 mg/kg
50 mg/kg
*** *** ***
Histamine mcg/ml
Figure 3. Effect of P. oleracea on histamine content of stomach homogenate of
rabbits. Values are represented as mean and n¼6, p<0.001, p<0.01
and p<0.05 as compare to control.
6 M. S. Z. JAN ET AL.
Gastric ulcer presents a major health problem having high
morbidity and mortality rate (Mejia and Kraft 2009). However,
the use of medicinal plants and their secondary metabolites
gain importance worldwide for the treatment of different dis-
eases. Literature reports revealed the gastro protective effect
of medicinal plants in different ulcer induced models in
rodents (Rahim et al. 2014).
The medicinal plants showed better antiulcer activity,
when compared to conventional agents like H
blockers, pro-
ton pump inhibitors and cytoprotective agents (Onasanwo
et al. 2011).
In the present study, antiulcer potential of crude extract
and subsequent fractions of P. oleracea was investigated as
part of our aims to establish the pharmacological potential of
this plant scientifically. Various folk uses have been reported
for this plant as described earlier, which include the gastro
protective activity. In this study, the mechanistic activity of
P. oleracea was also determined in animal models using acidi-
fied ethanol as ulcerogenic agent (Carlos et al. 2010).
Through its direct action, acidified ethanol penetrates the
gastric mucosa rapidly causing damage to gastrointestinal
mucosa cells and membranes (Kumar et al. 2011). The pro-
cess of formation of gastric mucosal lesions start through
micro vascular injury, increase of the vascular permeability,
which affected mucus secretion, pepsin secretion, and the
loss of hydrogen ions and histamine into the lumen (Szabo
et al. 1985). In this study, the consequential lesions of differ-
ent sizes along the glandular part of the stomach were devel-
oped by acidified ethanol. Our results showed that among
the fractions, EAF decreased ulcer index of rabbit which was
comparable to ranitidine. Histamine released by acidified
ethanol increases the production of HCl in the stomach
which decreases gastric pH (Dorababu et al. 2006). In the
present study EAF at 400 mg/kg reduced the acidity and
increased the pH of the gastric juice.
On the other hand, gastric mucus provides protection to
the gastric tissue against gastric acid. Our results showed
that EAF at 400 mg/kg significantly increased the mucus con-
tent against acidified ethanol (Allen and Flemstr
om 2005).
Pepsin also plays an important role in producing gastric
ulcer. It damages the junctional complex which make the
epithelial layer of stomach acid permeable, pepsinogen is
auto catalytically converted to pepsin at low pH. The pepsin
hydrolyze the collagen surface epithelium, especially type IV,
and damage mucosa. Therefore, inhibition of pepsin secre-
tion play important role in the management of gastric ulcer
(Selvamathy et al. 2010). Our results showed that EAF at
400 mg/kg significantly decreased pepsin content.
Moreover for the mechanistic study, three different path-
ways were determined. Histamine which is the main mediator
of gastric acid secretion, triggers a chain of events which
damage the mucosa (Mahmood et al. 2005). Histamine
increase gastric acid secretion via H
-receptors that are
coupled to G-protein system. Stimulation of adenyl cyclase
increases cAMP level, which is compulsory for activation of
protein kinases. The protein kinases cause the phosporylation
of certain phosphatases and activate the proton pump for
the production of HCl (Mesia-Vela et al. 2002). The oral
administration of P. oleracea extracts reduced the histamine
level which is the main pathway for blockade of gastric acid
secretion. Moreover, EAF at high dose (400 mg/kg, p.o.) inhib-
ited H
ATPase enzyme in comparison with positive control
group. The H
ATPase enzyme which is also known as pro-
ton pump, that exchange protons (H
) and potassium (K
ions across the plasma membrane of stomach and play vital
role in acid formation (Gumz et al. 2010). Similarly, acidified
ethanol can also augment the production of ROS i.e., super-
oxide anion and hydroxyl radicals (Bailey 2003). ROS react
with cellular lipids and form lipid peroxides. An effective indi-
cator of oxidative stress is MDA, the major metabolite of lipid
peroxidation (Rouhollahi et al. 2014). This study showed that
EAF at high dose (400 mg/kg, p.o.) prevented gastric ulcer-
ation by reducing MDA production against acidified ethanol.
This study explained that EAF of P. oleracea exhibit anti gas-
tric ulcerogenic effect through (i) decreasing histamine level
in tissue homogenate which is an important pathway of
decreasing gastric acidity, beside the solvent extracts also (ii)
decreased MDA levels in the tissue and (iii) moderately inhib-
ited H
ATPase. As the solvent extract inhibited gastric
ulcer via multiple pathways, so this study may provide an
important alternative, safe and effective remedy as compare
to available conventional treatment (PPIs and H
Isolation of active compounds from ethyl acetate fraction
responsible for anti gastric ulcer activity is our future plan.
100 mg/kg
200 mg /kg
400 mg /kg
10 mg/kg
10 mg/kg
nMole of phosphat e/min/mg protien
Figure 4. Effect of P. oleracea on H
ATPase activity in rabbits. Values are
represented as mean and n¼6, p<0.001, p<0.01 and p<0.05 as
compare to control.
Figure 5. Effect of P. oleracea on stomach tissue of rabbits (A) Micrograph of ranitidine þacidified ethanol (Positive control group) (B) Ulcer control group (C) EAF
(100 mg/kg þacidified ethanol (D) EAF 200 mg/kg þacidified ethanol (E) EAF 400 mg/kg) þacidified ethanol. ME: mucularis externa; M: Mucosa; SM: submucosa.
8 M. S. Z. JAN ET AL.
Disclosure statement
The authors declare that they have no competing interests.
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10 M. S. Z. JAN ET AL.
... In 2012, Ullah et al. reported lemongrass tea effectively reduced high blood pressure and decreased heart rate. 13 Koner et al. identified lemongrass tea as antioxidants to detoxify and clean the toxic substance within our body. It regulated blood circulation and lowered blood pressure by the potassium ion from lemongrass tea. ...
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Hypertension (high blood pressure) is the pre-symptom of car-diovascular disease. The number of people living with hyperten-sion has doubled to 1.28 billion and proportionally increased until today. This is a long-term disease and requires continuous monitoring. A traditional Chinese herbal, "Lemongrass", might be a good choice for the mainstay of hypertension. Some library search engines are used, such as SCI/SCIE, PubMed, and Scopus, within ten to twenty years, from 1999-2020. The searched keywords and phrases are "lemongrass", "formulation", "traditional Chinese medicine", "hydrogel", "hypertension", "lemongrass + tea formu-lation", "lemongrass + hydrogel", "Lemongrass + Hypertension", "Lemongrass + traditional Chinese medicine" etc. This mini-review discusses the background of hypertension, lemongrass, research progress, mechanism, lemongrass tea formulations, lemongrass with Traditional Chinese Medicine (TCM) formulations , and the lemongrass hydrogel application in the treatment of hypertension.
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Background: Melastoma malabathricum L. (family Melastomaceae; MM) and Muntingia calabura L. (family Elaeocarpaceae; MC) have been separately reported to possess gastroprotective activity. In an attempt to develop a pharmaceutical product with antiulcer potential, the synergistic gastroprotective activity of methanolic extract of a mixture of MM and MC (MMMC) at various ratios was evaluated in rat models. Methods: Rats were pre-treated orally with 2% Tween 80 (vehicle), 100 mg/kg ranitidine (reference drug) or MMMC (ratios of 1:1, 1:3 and 3:1 (v/v); doses of 15, 150 or 300 mg/kg) and then subjected to the ethanol-induced gastric ulcer or pyloric ligation assays. Stomach of rats from the former assay was collected and subjected to the macroscopic and microscopic observations, and enzymatic and non-enzymatic antioxidant studies while the gastric juice content and tissue from the latter assay were subjected to the antisecretory activity study. The UHPLC analysis of MMMC was also performed. Result: MMMC, in the ratio 1:1, demonstrated the most effective (P < 0.001) gastroprotective activity indicated by the highest reduction in ethanol-induced ulcer area formation. These macroscopic findings were supported by the microscopic observations. Except for pH and total acidity, MMMC also significantly (P < 0.001) reduced the volume of gastric content but increased the gastric wall mucus content in the pyloric-ligation test. MMMC also demonstrated remarkable antioxidant activity indicated by the highest total phenolic content (TPC) value and oxygen radical absorbance capacity (ORAC) activity with the recorded IC50 value of approximately 53 μg/mL for the 2,2- diphenyl-1-picrylhydrazyl (DPPH) scavenging activity. MMMC also improved the catalase (CAT), superoxide dismutase (SOD), glutathione (GSH), prostaglandin E2 (PGE2) and malondialdehyde (MDA) activities of the gastric tissue intoxicated by ethanol. UHPLC analysis of MMMC confirmed the presence several flavonoid-based bioactive compounds. Conclusion: MMMC, at the ratio of 1:1 (v/v), exerts gastroprotective activity partly by activating its antisecretory and antioxidant activities, and via modulation of the gastric tissue endogenous antioxidant system.
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Gastric ulcer is the most digestive disease found in clinical practice. Punicalagin (PCG) found in pomegranate juice has antioxidant and tissue repair properties. Therefore, the aim of this study was to investigate punicalagin's gastroprotective effect against ethanol-induce gastric injury. Four groups of rats; first group served as control, group 2: Treated with absolute ethanol (5 ml/kg, po), group 3: Rats were pre-treated with ranitidine (as a reference drug) (50 mg/kg, po) before ethanol and group 4: Pre-treated with PCG (4 mg/kg, po) before ethanol. Pre-treatment with PCG reduced ulcer index and histopathological changes, oxidative stress induced by ethanol while it elevated antioxidant activity. Although, it down regulated Tumor Necrosis Factor (TNF-α) gene expression and mucosal levels of inflammatory cytokines; TNF-α, Interleukin (IL-1β) and Interferon Gamma (IFNγ), it up regulated mucosal level of IL-10. Also, it reduced mucosal Nuclear Factor Kappa B (NFκB) protein expression, mylopeoxidase and caspase 3 activities as well as gene expression of caspase 9 while it elevated antiapoptotic B-Cell Lymphoma 2 (Bcl-2) gene expression, mucosal nitric oxide and mucin content. However, it had negative action on prostaglandin E2 and acid secretions. These findings indicate gastroprotective effects of punicalagin against ethanol induced gastric ulcer through suppression of mucosal oxidative stress and inflammation through NFκB pathway as well cytoprotective defences independent on prostaglandin E2 and acid secretions.
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Background Cibotium barometz is a medical herb used traditionally in the Malaysian peninsula for several ailments, including gastric ulcer. The aim of this study was assessment the anti-ulcer effects of C. barometz hair on ethanol-induced stomach hemorrhagic abrasions in animals. Seven groups of Sprague Dawley (SD) rats were administered 10% Tween 20 in the normal control and ulcer control groups, and omeprazole 20 mg/kg and 62.5, 125, 250, and 500 mg/kg of C. barometz hair extract in the experimental groups. After 60 min, the normal control group of rats was orally administered 10% Tween 20, while absolute ethanol was orally administered to the groups of ulcer control, omeprazole and experimental groups. Stomachs of the rats were examined macroscopically and histologically. Homogenates of stomachs were used to evaluate endogenous antioxidant enzyme activities. ResultsRats pre-fed with plant extract presented a significant decrease in the sore area, increased pH of gastric contents and preserved stomach wall mucus compared to the ulcer group. Histologically, rats pre-fed with C. barometz hair extract showed mild to moderate disruptions of the surface epithelium while animals pre-fed with absolute ethanol showed severe disruptions of the stomach epithelium with edema and leucocyte penetration of the submucosal layer. A Periodic acid Schiff (PAS) staining revealed that each rat pre-treated with the plant extract displayed an intense uptake of stomach epithelial glycoprotein magenta color compared to the ulcer control group. Immunohistochemical analysis revealed that rats pre-fed with the plant extract showed an up-regulation of the heat shock protein 70 (HSP70) and down-regulation of Bax proteins compared to ulcer control rats. Homogenates of the stomach tissue demonstrated significant increases in the endogenous antioxidant enzymatic activity and decreased lipid peroxidation (MDA) in rats pre-treated with C. barometz hair extract compared with the ulcer control rats. In acute toxicity, the liver and kidney revealed no hepatotoxic or nephrotoxic effects histologically. Conclusions The gastric cytoprotective action of C. barometz hair extract might be attributed to antioxidants, an increase in gastric pH, stomach mucus preservation, increased endogenous antioxidant enzymes, decreased lipid peroxidation, up-regulation of HSP70 and down-regulation of Bax proteins.
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Background Muntingia calabura L. (family Muntingiaceae), commonly known as Jamaican cherry or kerukup siam in Malaysia, is used traditionally to treat various ailments. The aim of this study is to elucidate the possible underlying gastroprotective mechanisms of ethyl acetate fraction (EAF) of Muntingia calabura methanolic leaves extract (MEMC). Methods MEMC and its fractions were subjected to HPLC analysis to identify and quantify the presence of its phyto-constituents. The mechanism of gastroptotection of EAF was further investigated using pylorus ligation-induced gastric lesion rat model (100, 250, and 500 mg/kg). Macroscopic analysis of the stomach, evaluation of gastric content parameters such as volume, pH, free and total acidity, protein estimation, and quantification of mucus were carried out. The participation of nitric oxide (NO) and sulfhydryl (SH) compounds was evaluated and the superoxide dismutase (SOD), gluthathione (GSH), catalase (CAT), malondialdehyde (MDA), prostaglandin E2 (PGE2) and NO level in the ethanol induced stomach tissue homogenate was determined. Results HPLC analysis confirmed the presence of quercetin and gallic acid in EAF. In pylorus-ligation model, EAF significantly (p <0.001) prevent gastric lesion formation. Volume of gastric content and total protein content reduced significantly (p < 0.01 and p < 0.05, respectively), while free and total acidity reduced in the doses of 250 and 500 mg/kg (p <0.001 and p <0.05, respectively). EAF also augmented the mucus content significantly (p < 0.001). Pre-treatment with N-nitro-L-arginine methyl ester (L-NAME) or N-ethylmaleimide (NEM) reversed the gastroprotective activity of EAF. EAF treatment markedly ameliorated the SOD, GSH and CAT activity and PGE2 and NO level while attenuating MDA level, relative to the vehicle group. Conclusions In conclusion, the underlying gastroprotective mechanisms of EAF could be associated with the antisecretory, participation of mucus, antiperoxidative, improvement of antioxidant status, modulation of NO and SH compounds, stimulation of PGE2 as well as presence of quercetin and gallic acid.
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Context: Muntingia calabura L. (family Muntingiaceae) and Melastoma malabathricum L. (family Melastomaceae) are traditionally used to treat gastric ulcer. Objective: The present study determines the mechanisms of gastroprotective activity of the chloroform extract of leaves obtained from both the plants using several in vitro and in vivo assays. Materials and methods: Phytochemical screening, HPLC analysis, and antioxidant activity of the respective extract were carried out. Gastroprotective activity was determined using ethanol-induced gastric ulcer assay while the mechanisms of gastroprotection were determined using the pyloric ligation assay. The test solutions [8% Tween-80 (vehicle), 20 mg/kg omeprazole, and different doses of extracts (50, 250, or 500 mg/kg] were administered orally once daily for 7 consecutive days before the animals were subjected to ethanol induced gastric ulcers. Results: The chloroform-extracted M. calabura (CEMC) contains tannins, polyphenolics, triterpenes, and steroids while the chloroform-extracted M. malabathricum (CEMM) contains only triterpenes and steroids. CEMC, but not CEMM, exerted remarkably strong antioxidant activity in the 2,2-diphenyl-1-picrylhydrazyl (DPPH)- (86% versus 16%) and superoxide- (73% versus 36%) radical scavenging assays. Both extracts demonstrated significant (p < 0.05) gastroprotection with the EC50 value recorded at 192.3 or 297.7 mg/kg, respectively. In the pylorus ligation assay, CEMC and CEMM significantly (p < 0.05) reduced the total and free acidity and volume; while increased the pH of gastric juice as well as the gastric wall mucus content in comparison with the vehicle-treated group. Discussion and conclusion: CEMC and CEMM exert gastroprotective effects in animals with ethanol-induced gastric ulcers via antioxidant and anti-secretory effects.
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Gastric peptic ulcer is one of the common disorders of gastrointestinal tract, which occur due to an imbalance between the offensive and defensive factors. It is an illness that affects a considerable number of people worldwide. This study was conducted to evaluate the antiulcerogenic and antiulcer effects and recognize the basic mechanism of action of Piptadeniastrum africanum stem bark extracts. The aqueous and methanol extracts of Piptadeniastrum africanum were administered at the doses 125, 250 and 500 mg/kg to evaluate their effects on gastric ulcer induced by the HCl/ethanol mixture, indomethacin and acetic acid in Wistar strain male adult rats, aged between 12 and 16 weeks and weighing between 180 and 220 g. Ranitidine, Maalox and Misoprostol were used as standard drugs. Histopathological examination and nitric oxide level were performed to evaluate the basic mechanism of action of Piptadeniastrum africanum. Phytochemical screening was carried out to identify known phytochemicals present in these extracts. The aqueous and methanol extracts of stem bark of Piptadeniastrum africanum significantly inhibited (p < 0.01) gastric ulceration induced by HCl/ethanol to the percentages of inhibition of 81.38; 98.75 and 100 % for the aqueous extract and then 75.83, 89.76 and 96.52 % for the methanol extract, and with the Indomethacin-induced ulcers, aqueous and methanol extracts of bark of Piptadeniastrum africanum reduce significantly (p < 0.01) induced gastric lesions in rats, with percentage of cure 35.75; 52.33 and 98.58 % for the aqueous extract, and 33.7; 51.97; and 65.93 to the methanol extract. The results revealed a significant reduction of ulcerated surface in both extracts and increase of nitric oxide (NO) level with methanol extract. When compared to methanol extract, aqueous extract showed more pronounced effects, corresponding to percentages of healing of 59. 92; 84.12 and 59.65 % for the aqueous extract; and 70.43; 55.49 and 57.59 % for the methanol extract in the ulcer induced by acetic acid, all at the respective doses of 125, 250 and 500 mg/kg. Histopathological observations also demonstrated curative effect. As such, both extracts were found to exhibit preventive and curative effects through the release of NO and growth factors. This could also be due to the presence of phytochemicals such as alkaloids, flavonoids, phenols and saponins which act as antisecretory agents. Piptadeniastrum africanum stem bark extracts thus have gastroprotective and ulcer healing effects, which could result from their activities by stimulating important cellular mechanisms such as migration and proliferation of epithelial cells that may have a cytoprotective effect by stimulating the release of prostaglandins. These results are required to confirm the ethnopharmacological use of Piptadeniastrum africanum stem bark in the treatment of ulcer.
The aim of the present study was to evaluate the gastroprotective effect of hydro alcoholic extract of Swertia chirayita (SCHAE) on experimental ulcers in rats.Male albino Wistar rats were induced for gastric ulcer by ethanol, pyloric ligation (PL) and cold restraint stress (CRS) after pretreatment with SCHAE (100/200mg/ kg b.wt) for 30 days. The gastro protective effect of SCHAE was assessed by lesion index, pH, titrable acidity, level of pepsin, lipid peroxides (LPO), reduced glutathione (GSH), myeloperoxidase activity and by the levels of enzymatic antioxidants superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx), nucleic acid and protein bound carbohydrates. The gastric tissues were used for the determination of adherent mucus content and also for the histological examination. A significant reduction in lesion index were observed in ulcer induced animals pretreated with 200mg/kg b.wt of SCHAE. The mucin content was found to be restored significantly in ulcer induced SCHAE pre-treated animals. The alterations observed in the level of LPO, pH, titrable acidity were found to be minimized in SCHAE treated animals. Histological observations on gastric mucosa also confirmed the gastro protective activity of SCHAE. Normal rats treated with SCHAE did not show any toxicity. From the results of this study, it could be concluded that SCHAE acts as an antiulcer agent probably by its free radical scavenging activity and mucin protective nature.
AIM: To illustrate the pathophysiological role of metallothionein (MT) in gastric ulcer induced by stress. METHODS: Wistar rats underwent water-immersion-restraint (WIR) stress, ZnSO4 (an MT inducer) treatment, WIR+ZnSO4 or WIR+MT, and the ulcer index (UI) was estimated in excised stomach and liver tissues. The mRNA level of gastric MT was determined by semi-quantitative RT-PCR. The MT content in gastric and hepatic tissues was determined by Cd/hemoglobin affinity assay. The lipid peroxidation products malondialdehyde (MDA) and conjugated dienes (CD) were estimated by use of thiobarbituric acid reactive species and ultraviolet spectrophotometry. RESULTS: WIR stress induced severe gastric mucosal lesions in rats. Compared with control rats, stressed rats had increased lipid peroxide content in serum and stomach and liver tissues. MDA content was increased by 34%, 21% and 29% and CD level by 270%, 83% and 28%, respectively. MT content in the stomach and liver was increased by 0.74- and 1.8-fold, and the MT-mRNA level in the stomach was increased by 26%. Pretreatment with ZnSO4 prevented gastric lesion development (the UI was 87% lower than that without pretreatment), and the MDA and CD content in serum and tissues was lower. The MT content in the liver was double in rats that were not pretreated, and the MT mRNA level in the stomach was 35% higher. MT administration 1 h before the WIR stress prevented gastric lesion development (the UI decreased by 47% compared with that in rats not pretreated), and the MDA and CD content in serum and tissues was significantly lower. CONCLUSION: In WIR-stressed rats, the MT level was increased in serum and in stomach and liver tissues. Pre-administration of exogenous MT or pre-induction of endogenous MT can protect the gastric mucosa against stress-induced ulcers and inhibits the formation of stress-induced lipid peroxide. MT could have a gastroprotective effect and might be a new interventive and therapeutic target in stress-induced gastric ulcers.