BACKGROUND
peptic ulcer disease  is a common condition as contributes to morbidity and mortality in humans worldwide.
the annual prevalence of pud is  <dig> - <dig> %  <cit> .
aetiology of pud is multi-factorial  <cit> .
high acid secretion and decreased mucosal barrier contribute to initiation and progression of pud  <cit> .
the aetiology is ranging from helicobacter pylori  infection to stress, lifestyle characteristics and drugs  <cit> .
one of the most important drugs that contribute to pud is non-steroidal anti-inflammatory drugs   <cit> .
consumption of nsaids is shown to be related to increased mucosal injury, erosions and gastric and duodenal bleeding.
this effect is mainly due to the inhibition of the cyclooxygenase  <dig>  resulting in reduced mucosal defence as well as thromboxane a <dig> which regulates platelet aggregation and therefore results in gastric bleeding  <cit> .
another mechanism that is well described in nsaids induced gastric injury is the production of reactive oxygen species   <cit> .
in literatures, several authors had been reported the use of synthesized compound for the treatment of gastrointestinal disorders including gastric ulcerogenic damage  <cit> .
the main goals of pud treatment, is firstly to reduce acid secretion, the hallmark of these medications is the proton pomp inhibitor , and secondly to improve mucosal barrier, including sucralfate and bismuth salts   <cit> .
it is also believed that scavenging free radicals will also prevent the production of pud  <cit> .
after discovery of h-pylori, eradication of this bacterial infection became one of the essential steps in treatment of pud  <cit> .
herbal medication and natural substances were long used for the treatment and management of pud.
different herbs and spices have been introduced, in traditional medicines, as treatments for pud.
plant derived medications have long been considered as safe and effective agents against pud  <cit> .
flavonoids are one of the most abundant compartments in the vegetarian kingdom.
flavonoids are found in any part of plants including leaves, stalks, roots, fruits and seeds and are proven to have different effects on the gastrointestinal tract.
they are also shown to be effective in prevention and treatment of pud  <cit> .
chalcones are the precursors of flavonoids in plants.
most chalcones contain a six-member heterocyclic ring and are considered the first product of flavonoid synthesis pathway  <cit> .
various chalcones have been identified in nature and synthesized in laboratories and were shown to have variety of effects including prevention of peptic ulcer and oxidative stress as well as antibiotic, antifungal and anti-inflammatory effects  <cit> .
among the variety of chalcones  <dig> chalcones were shown to have antiulcer effects due to increased gastric blood flow and stimulating mucosal secretion.
to the best of our knowledge no study was performed on the protective effect of chalcone against ulcers induced by nsaid prescription as well as the antioxidant properties of this chalcone in nsaid induced gastric injury.
the aim of this study was to identify the effects of chalcone -3-m-tolyl-propenone)  on nsaid induced erosive gastic damage in sprague dawley rats.
RESULTS
acute toxicity
a single oral administration of -3-m-tolyl-propenone)  chalcone at three doses  did not result any mortality for 24 hours and no toxic effects and abnormal behaviour  were observed throughout 14 days.
the biochemical measurement of blood serum, clinical observation and histo-pathological estimation of the kidney and liver revealed that there are no significant differences  between treated groups and normal control group as shown in , .table  <dig>
effects of hptp on renal function test  in acute toxicity studies
male
sodium mmol/l
potassium mmol/l
chloride mmol/l
co
2
mmol/l
an.gap mmol/l
urea mmol/l
creatinine μmol/l
female
values are expressed as mean ± sem and one-way anova was used for the analysis.
effects of hptp on liver functions in rats  in acute toxicity studies
animals
t. protein
albumin
globulin
t. billirubin
alp
alt
ast
male
female
values are expressed as mean ± sem and one-way anova was used for the analysis.
effects of hptp on lipid profile levels in blood serum of male and female rats in acute toxicity studies
animals
triglyceride
t. cholesterol
hdl
male
female
values are expressed as mean ± sem and one-way anova was used for the analysis.
effects of hptp on histology of liver and kidney in acute toxicity testing.
rats treated with vehicle , rats treated with 250 mg/kg hptp , rats treated with 500 mg/kg hptp , rats treated with 1000 mg/kg hptp .
there were no significant difference in the histology of liver and kidney between treated and control groups  .
antiulcer characteristics
the gastro-protective effects of hptp chalcone was estimated against indomethacin induced gastric ulcer in rats  in terms of ph of gastric, mucus barrier, erosive gastric damage area and percentage of inhibition.
pre-treatment with 50 mg/kg and 100 mg/kg of hptp were found to inhibit stomach lining injury induced by indomethacin.
this inhibition appeared in low dose and high dose  respectively.
thus, this study revealed that hptp can decrease the acidity significantly  which plays a major role in stimulating stomach ulcer.
gastric mucus layer that is covering the gastric lining plays an important role in preserving the inner layer of stomach from exogenous aggressive factors.
in this study it has been shown that hptp significantly enhanced the gastric mucus production at low dose and high dose administration .table  <dig>
effects of hptp on gastric ulcer induced by indomethacin in rats
pre-treatment
gastric ph
mucus barrier μg/g tissue
ulcer area
% inhibition
values are expressed as mean ± sem.
indicates significant at p <  <dig>  versus indomethacin  group with pre-treatment groups.
the macroscopic sections of stomachs showed significant differences between groups .
rats in the hptp treated groups  significantly reduced areas of gastric lesions compared to rats in group  <dig>  indomethacin induced gastric lesions were significantly reduced in term of size and severity in rats pre-treated with omeprazole as shown in figure  <dig> figure  <dig>
macroscopic evaluation of the gastric lesions in rats.
the normal control group  shows no injury of gastric mucosa.
the indomethacin treated group  produced visible hemorrhagic necrosis of gastric mucosa .
rats pre-treated with 50 mg/kg hptp  shows moderate injuries in the gastric mucosa  and little flattened .
rats pretreated with 100 mg/kg hptp  shows little flattened  in gastric mucosa and fewer injuries were observed .
the omeprazole treated group  has more flattened  of gastric mucosa and few and small injuries in the gastric mucosa .
antioxidant properties
the enzymatic and non-enzymatic parameters that play important role in protecting the gastric mucosa from damage.
this study evaluated the effect of ulcer induction on some of these parameters , thus the effect of hptp pre-treatment on these parameters production.
it has been found that in indomethacin group gpx, sod, and pge <dig> were significantly lower than in normal control , while mda was significantly higher in indomethacin group than in control .
this indicated that indomethacin increase the level of mda and decrease the level of sod, gpx and pge <dig>  as for hptp, this study revealed that chalcone significantly increased sod and pge <dig> at low and high dose, while hptp chalcone increased gpx level but no significant differences between indomethacin group and hptp groups.
in this study it has been shown that mda level in indomethacin group was increased but hptp chalcone at low and high doses decreased mda that play important role in gastric ulcerogenic effects .figure  <dig>
effects of hptp on glutathione peroxidase activity in indomethacin induced erosive gastric damage in rat stomach.
values are expressed as mean ± s.e.m.
indicates significance at p <  <dig>  versus indomethacin with hptp chalcone and  at p <  <dig>  versus indomethacin with normal control.
effect of hptp on sod enzyme in indomethacin induced erosive gastric damage in rat stomach.
values are expressed as mean ± sem.
indicates significance at p <  <dig>  versus indomethacin with hptpchalcone and  at p <  <dig>  versus indomethacin with normal control.
effect of hptp on prostaglandin e
2
activity.
values are expressed as mean ± s.e.m.
indicates significance at p <  <dig>  versus indomethacin with hptp chalcone and  at p <  <dig>  versus indomethacin with normal control.
effect of pre-treatment with hptp and omeprazole followed by indomethacin  on malondialdehyde releasing.
values are expressed as mean ± s.e.m.
indicates significance at p <  <dig>  versus indomethacin with normal control and  at p <  <dig>  versus hptp chalcone.
sod, gpx, pge
2
and lipid peroxidation properties of hptp in indomethacin-induced gastric lesions in rats
pre-treatment
gpx
sod
pge <dig>
mda
values are expressed as mean ± sem.
indicates significance at p <  <dig>  versus normal control with indomethacin group,  p <  <dig>  versus indomethacin with all treated groups.
gpx = glutathione peroxidase, sod = superoxide dismutase, pge2 = prostaglandin e <dig>  mda = estimation of lipid peroxidation level.
histological examination
histological examinations of group  <dig> indicated that there was no disruption of the surface epithelium, while the histological examination showed extensive damage to the gastric mucosa in group  <dig>  with necrotic lesions penetrating deeply into the mucosa accompanied by extensive edema and leukocyte infiltration of the submucosal layer .
group  <dig> exhibited moderate disruption of the surface epithelium, with edema and leukocyte infiltration of the submucosal layer, and group  <dig> showed a mild disruption of the surface epithelium with edema and leukocyte infiltration into the submucosal layer.
group  <dig> showed mild edema and leukocyte infiltration of the submucosal layer, but no disruption of the surface epithelium.
these results demonstrated that the hptp exerted cytoprotective effects in a dose-dependent manner .figure  <dig>
histological evaluation of gastric lesions in sections stained with hematoxylin & eosin .
normal control group  has normal tissue epithelium.
indomethacin group  shows disruption of surface epithelium with hemorrhage  and the lesions infiltrate deep into mucosa layer  wide edema  and leukocyte infiltration .
hptp pre-treated groups  shows reduction of submucosal edema , mild to moderate disruption of mucosa epithelium  few infiltration of leukocyte.
omeprazole group  shows mild disruption of the surface epithelium , and mild submucosal edema  and bit of leukocyte .
shows the periodic acid-schiff  stain.
the gastric mucosa in animals pretreated with hptp or omeprazole  displayed increased pas staining intensity compared to the rats in group  <dig>  indicating an increase in the glycoprotein content of gastric mucosa in pretreated rats .figure  <dig>
histological evaluation of gastric glycoproteins in sections stained with pas.
the magenta color in the apical epithelial cells shows glycoprotein accumulation in the gastric glands .
normal control group , indomethacin treated group , hptp pre-treated groups  and omeprazole pre-treated group  .
the expression of the hsp <dig> and tgfβ protein in the gastric mucosa was down-regulated in section of ulcerated stomach tissue while it was up regulated in the groups pre-treated with  <dig>  100 mg/kg of hptp and 20 mg/kg of omeprazole respectively .figure  <dig>
immunohistochmical analysis of hsp <dig> in gastric tissue induced with indomethacin.
section showed normal mucosal.
section of ulcerated stomach tissue showed down-regulation of hsp <dig> in injured area .
sections  that pre-treated with  <dig>  100 mg/kg of hptp and 20 mg/kg of omeprazole respectively showed up-regulation of hsp <dig>  .
immunohistochmical analysis of tgf-β protein expression in gastric tissue of rat induced with indomethacin.
section shows normal mucosal area and express regulated of tgf-β .
section of ulcerated stomach tissue ashowed down-regulation of tgf-β almost non-existent in injured area .
sections  that pre-treated with  <dig>  100 mg/kg of hptp and 20 mg/kg of omeprazole respectively found the up-regulation of tgf-β  .
immunohistochmical analysis of bax protein expression in gastric tissue of ulcerated stomach showed up regulation of bax protein in injured area while it was down regulated in groups pre-treated with  <dig>  100 mg/kg of hptp and 20 mg/kg of omeprazole respectively .figure  <dig>
immunohistochmical analysis of bax protein expression in gastric tissue of rat induced with indomethacin.
section shows normal mucosal area and express down-regulated of bax.
section of ulcerated stomach tissue appears up-regulation of bax protein in injured area .
sections  that pre-treated with  <dig>  100 mg/kg of hptp and 20 mg/kg of omeprazole respectively found the bax protein expression was down-regulated .
discussion
chalcones are widely found in the plants and produce antioxidant, antimicrobial, anticancer and antibiotic properties.
their availability in different plant sources as well as their pharmacological values and low toxicity resulted in the interest of scientists and industries  <cit> .
this study was performed with the aim of evaluating antiulcer effects of tested chalcone in rats.
the results of this study revealed that all doses  of -3-m-tolyl-propenone)  chalcone are safe because none of the animals produced toxic signs and symptoms and none died during the study period.
moreover there are no significant differences in biochemical parameters of liver and kidney.
histology revealed no nephrotoxic or hepatotoxic effect.
similarly, several researcher used various synthesized schiff base compound and showed no nephrotoxicity or hepatotoxicity and the biochemical parameter are within the normal rang  <cit> .
based on these findings the doses of chalcone used for the antiulcer experiment was of no toxicity to rats.
moreover, this study showed that chalcone was able to produce more ulcer inhibition in gastric tissue in rats  chain process  <cit> .
odabasoglu et al.
also reported that indomethacin administration resulted in significant reduction in sod levels in rats.
this was in line with findings of this study that revealed a significant decrease in sod levels in rats with indomethacin induced gastric ulcer.
this study also revealed that administration of chalcone is capable of increasing the sod levels and therefore play a preventive role against indomethacin-induced gastric ulcerogenic damage.
koizumi et al.
<cit>  showed that the administration of 30 mg/kg indomethacin resulted in a significant reduction in the pge <dig> levels in rats.
this finding was also in line with the findings of this study.
this study revealed that the most important oxidative mechanisms of indomethacin induced erosive gastric damage is related to reduction of the important antioxidants in the stomach including sod as well as pge <dig>  in the present study, the findings from histology tests showed the multiple potential effect of chalcone through reduction in acidity and inhibition percentage, while in the same time mucus barrier was increased, which resulted in smaller gastric ulcer area in chalcone groups .
these properties were also found to be dose dependent as they improved gastric mucosal state by increasing dosage.
furthermore, this study revealed that administration of chalcone at low and high dose  reduced ulcers in stomach.
these findings were in line with the previous findings in terms of protective mechanisms of chalcone and flavonoids  <cit> .
the findings of this study suggest the use of chalcone with nsaids to reduce the risk of pud.
also in a study by okunrobo and colleagues  <cit>  they tested the anti-inflammatory and gastro-protective properties of a synthetic chalcone with different dosages on rats who compared with the acetylsalicylic acid  and cimetidine  rat groups.
the results were similar with the results present study terms of gastric wall mucosal characteristics  and acidity of gastric content.
finally omeprazole was used as antiulcer reference chemicals in this study against induced ulcer by indomethacin.
omeprazole as a proton pump inhibitors  are broadly prescribed medications with the control of gastric acid secretion for the prevention or treatment of peptic ulcer  <cit> .
omeprazole can suppress the gastric acid secretion through the h+,k + −atpase  <cit> .
it has been discovered that the h+,k + −atpase as gastric cell proton pump transfers h+ in exchange for luminal k+ to generate a highly acidic environment in the stomach that is responsible for gastric acid secretion  <cit> .
lastly indomethacin as a potent inhibitor of prostaglandins synthesis diminishes the protective effect of pge <dig> on mucosa of gastric wall  <cit> .
CONCLUSIONS
acute toxicity tests did not show any sign of morbidity or mortality up to 1000 mg/kg of hptp.
rats pre-treated with 50 mg/kg or 100 mg/kg of hptp before administration of 100 mg/kg of indomethacin significantly protect erosive gastric mucosal damage compared to ulcerogenic control group.
histologic findings showed that hptp remarkably protect gastric mucosa compared to ulcerogenic control group.
the gastroprotective effects of hptp could be attributed to increase in ph and mucus content of gastric content, increase endogenous enzymes and pge <dig>  and decrease in mda level in gastric homogenate, over-expresion of hsp <dig> and tgf-β proteins, and down-expression of bax protein.

 peptic ulcer disease is a common condition as contributes to morbidity and mortality in humans worldwide. the annual prevalence of pud is - %. aetiology of pud is multi-factorial. high acid secretion and decreased mucosal barrier contribute to initiation and progression of pud. the aetiology is ranging from helicobacter pylori infection to stress, lifestyle characteristics and drugs. one of the most important drugs that contribute to pud is non-steroidal anti-inflammatory drugs. consumption of nsaids is shown to be related to increased mucosal injury, erosions and gastric and duodenal bleeding. this effect is mainly due to the inhibition of the cyclooxygenase resulting in reduced mucosal defence as well as thromboxane a which regulates platelet aggregation and therefore results in gastric bleeding. another mechanism that is well described in nsaids induced gastric injury is the production of reactive oxygen species. in literatures, several authors had been reported the use of synthesized compound for the treatment of gastrointestinal disorders including gastric ulcerogenic damage. the main goals of pud treatment, is firstly to reduce acid secretion, the hallmark of these medications is the proton pomp inhibitor, and secondly to improve mucosal barrier, including sucralfate and bismuth salts. it is also believed that scavenging free radicals will also prevent the production of pud. after discovery of h-pylori, eradication of this bacterial infection became one of the essential steps in treatment of pud. herbal medication and natural substances were long used for the treatment and management of pud. different herbs and spices have been introduced, in traditional medicines, as treatments for pud. plant derived medications have long been considered as safe and effective agents against pud. flavonoids are one of the most abundant compartments in the vegetarian kingdom. flavonoids are found in any part of plants including leaves, stalks, roots, fruits and seeds and are proven to have different effects on the gastrointestinal tract. they are also shown to be effective in prevention and treatment of pud. chalcones are the precursors of flavonoids in plants. most chalcones contain a six-member heterocyclic ring and are considered the first product of flavonoid synthesis pathway. various chalcones have been identified in nature and synthesized in laboratories and were shown to have variety of effects including prevention of peptic ulcer and oxidative stress as well as antibiotic, antifungal and anti-inflammatory effects. among the variety of chalcones chalcones were shown to have antiulcer effects due to increased gastric blood flow and stimulating mucosal secretion. to the best of our knowledge no study was performed on the protective effect of chalcone against ulcers induced by nsaid prescription as well as the antioxidant properties of this chalcone in nsaid induced gastric injury. the aim of this study was to identify the effects of chalcone -3-m-tolyl-propenone) on nsaid induced erosive gastic damage in sprague dawley rats.  acute toxicity a single oral administration of -3-m-tolyl-propenone) chalcone at three doses did not result any mortality for 24 hours and no toxic effects and abnormal behaviour were observed throughout 14 days. the biochemical measurement of blood serum, clinical observation and histo-pathological estimation of the kidney and liver revealed that there are no significant differences between treated groups and normal control group as shown in,.table effects of hptp on renal function test in acute toxicity studies male sodium mmol/l potassium mmol/l chloride mmol/l co 2 mmol/l an.gap mmol/l urea mmol/l creatinine μmol/l female values are expressed as mean ± sem and one-way anova was used for the analysis. effects of hptp on liver functions in rats in acute toxicity studies animals t. protein albumin globulin t. billirubin alp alt ast male female values are expressed as mean ± sem and one-way anova was used for the analysis. effects of hptp on lipid profile levels in blood serum of male and female rats in acute toxicity studies animals triglyceride t. cholesterol hdl male female values are expressed as mean ± sem and one-way anova was used for the analysis. effects of hptp on histology of liver and kidney in acute toxicity testing. rats treated with vehicle, rats treated with 250 mg/kg hptp, rats treated with 500 mg/kg hptp, rats treated with 1000 mg/kg hptp. there were no significant difference in the histology of liver and kidney between treated and control groups. antiulcer characteristics the gastro-protective effects of hptp chalcone was estimated against indomethacin induced gastric ulcer in rats in terms of ph of gastric, mucus barrier, erosive gastric damage area and percentage of inhibition. pre-treatment with 50 mg/kg and 100 mg/kg of hptp were found to inhibit stomach lining injury induced by indomethacin. this inhibition appeared in low dose and high dose respectively. thus, this study revealed that hptp can decrease the acidity significantly which plays a major role in stimulating stomach ulcer. gastric mucus layer that is covering the gastric lining plays an important role in preserving the inner layer of stomach from exogenous aggressive factors. in this study it has been shown that hptp significantly enhanced the gastric mucus production at low dose and high dose administration.table effects of hptp on gastric ulcer induced by indomethacin in rats pre-treatment gastric ph mucus barrier μg/g tissue ulcer area % inhibition values are expressed as mean ± sem. indicates significant at p < versus indomethacin group with pre-treatment groups. the macroscopic sections of stomachs showed significant differences between groups. rats in the hptp treated groups significantly reduced areas of gastric lesions compared to rats in group indomethacin induced gastric lesions were significantly reduced in term of size and severity in rats pre-treated with omeprazole as shown in figure  figure macroscopic evaluation of the gastric lesions in rats. the normal control group shows no injury of gastric mucosa. the indomethacin treated group produced visible hemorrhagic necrosis of gastric mucosa. rats pre-treated with 50 mg/kg hptp shows moderate injuries in the gastric mucosa and little flattened. rats pretreated with 100 mg/kg hptp shows little flattened in gastric mucosa and fewer injuries were observed. the omeprazole treated group has more flattened of gastric mucosa and few and small injuries in the gastric mucosa. antioxidant properties the enzymatic and non-enzymatic parameters that play important role in protecting the gastric mucosa from damage. this study evaluated the effect of ulcer induction on some of these parameters, thus the effect of hptp pre-treatment on these parameters production. it has been found that in indomethacin group gpx, sod, and pge were significantly lower than in normal control, while mda was significantly higher in indomethacin group than in control. this indicated that indomethacin increase the level of mda and decrease the level of sod, gpx and pge as for hptp, this study revealed that chalcone significantly increased sod and pge at low and high dose, while hptp chalcone increased gpx level but no significant differences between indomethacin group and hptp groups. in this study it has been shown that mda level in indomethacin group was increased but hptp chalcone at low and high doses decreased mda that play important role in gastric ulcerogenic effects.figure effects of hptp on glutathione peroxidase activity in indomethacin induced erosive gastric damage in rat stomach. values are expressed as mean ± s.e.m. indicates significance at p < versus indomethacin with hptp chalcone and at p < versus indomethacin with normal control. effect of hptp on sod enzyme in indomethacin induced erosive gastric damage in rat stomach. values are expressed as mean ± sem. indicates significance at p < versus indomethacin with hptpchalcone and at p < versus indomethacin with normal control. effect of hptp on prostaglandin e 2 activity. values are expressed as mean ± s.e.m. indicates significance at p < versus indomethacin with hptp chalcone and at p < versus indomethacin with normal control. effect of pre-treatment with hptp and omeprazole followed by indomethacin on malondialdehyde releasing. values are expressed as mean ± s.e.m. indicates significance at p < versus indomethacin with normal control and at p < versus hptp chalcone. sod, gpx, pge 2 and lipid peroxidation properties of hptp in indomethacin-induced gastric lesions in rats pre-treatment gpx sod pge mda values are expressed as mean ± sem. indicates significance at p < versus normal control with indomethacin group, p < versus indomethacin with all treated groups. gpx = glutathione peroxidase, sod = superoxide dismutase, pge2 = prostaglandin e mda = estimation of lipid peroxidation level. histological examination histological examinations of group indicated that there was no disruption of the surface epithelium, while the histological examination showed extensive damage to the gastric mucosa in group with necrotic lesions penetrating deeply into the mucosa accompanied by extensive edema and leukocyte infiltration of the submucosal layer. group exhibited moderate disruption of the surface epithelium, with edema and leukocyte infiltration of the submucosal layer, and group showed a mild disruption of the surface epithelium with edema and leukocyte infiltration into the submucosal layer. group showed mild edema and leukocyte infiltration of the submucosal layer, but no disruption of the surface epithelium. these results demonstrated that the hptp exerted cytoprotective effects in a dose-dependent manner.figure histological evaluation of gastric lesions in sections stained with hematoxylin & eosin. normal control group has normal tissue epithelium. indomethacin group shows disruption of surface epithelium with hemorrhage and the lesions infiltrate deep into mucosa layer wide edema and leukocyte infiltration. hptp pre-treated groups shows reduction of submucosal edema, mild to moderate disruption of mucosa epithelium few infiltration of leukocyte. omeprazole group shows mild disruption of the surface epithelium, and mild submucosal edema and bit of leukocyte. shows the periodic acid-schiff stain. the gastric mucosa in animals pretreated with hptp or omeprazole displayed increased pas staining intensity compared to the rats in group indicating an increase in the glycoprotein content of gastric mucosa in pretreated rats.figure histological evaluation of gastric glycoproteins in sections stained with pas. the magenta color in the apical epithelial cells shows glycoprotein accumulation in the gastric glands. normal control group, indomethacin treated group, hptp pre-treated groups and omeprazole pre-treated group. the expression of the hsp and tgfβ protein in the gastric mucosa was down-regulated in section of ulcerated stomach tissue while it was up regulated in the groups pre-treated with 100 mg/kg of hptp and 20 mg/kg of omeprazole respectively.figure immunohistochmical analysis of hsp in gastric tissue induced with indomethacin. section showed normal mucosal. section of ulcerated stomach tissue showed down-regulation of hsp in injured area. sections that pre-treated with 100 mg/kg of hptp and 20 mg/kg of omeprazole respectively showed up-regulation of hsp. immunohistochmical analysis of tgf-β protein expression in gastric tissue of rat induced with indomethacin. section shows normal mucosal area and express regulated of tgf-β. section of ulcerated stomach tissue ashowed down-regulation of tgf-β almost non-existent in injured area. sections that pre-treated with 100 mg/kg of hptp and 20 mg/kg of omeprazole respectively found the up-regulation of tgf-β. immunohistochmical analysis of bax protein expression in gastric tissue of ulcerated stomach showed up regulation of bax protein in injured area while it was down regulated in groups pre-treated with 100 mg/kg of hptp and 20 mg/kg of omeprazole respectively.figure immunohistochmical analysis of bax protein expression in gastric tissue of rat induced with indomethacin. section shows normal mucosal area and express down-regulated of bax. section of ulcerated stomach tissue appears up-regulation of bax protein in injured area. sections that pre-treated with 100 mg/kg of hptp and 20 mg/kg of omeprazole respectively found the bax protein expression was down-regulated. discussion chalcones are widely found in the plants and produce antioxidant, antimicrobial, anticancer and antibiotic properties. their availability in different plant sources as well as their pharmacological values and low toxicity resulted in the interest of scientists and industries. this study was performed with the aim of evaluating antiulcer effects of tested chalcone in rats. the results of this study revealed that all doses of -3-m-tolyl-propenone) chalcone are safe because none of the animals produced toxic signs and symptoms and none died during the study period. moreover there are no significant differences in biochemical parameters of liver and kidney. histology revealed no nephrotoxic or hepatotoxic effect. similarly, several researcher used various synthesized schiff base compound and showed no nephrotoxicity or hepatotoxicity and the biochemical parameter are within the normal rang. based on these findings the doses of chalcone used for the antiulcer experiment was of no toxicity to rats. moreover, this study showed that chalcone was able to produce more ulcer inhibition in gastric tissue in rats chain process. odabasoglu et al. also reported that indomethacin administration resulted in significant reduction in sod levels in rats. this was in line with findings of this study that revealed a significant decrease in sod levels in rats with indomethacin induced gastric ulcer. this study also revealed that administration of chalcone is capable of increasing the sod levels and therefore play a preventive role against indomethacin-induced gastric ulcerogenic damage. koizumi et al. showed that the administration of 30 mg/kg indomethacin resulted in a significant reduction in the pge levels in rats. this finding was also in line with the findings of this study. this study revealed that the most important oxidative mechanisms of indomethacin induced erosive gastric damage is related to reduction of the important antioxidants in the stomach including sod as well as pge in the present study, the findings from histology tests showed the multiple potential effect of chalcone through reduction in acidity and inhibition percentage, while in the same time mucus barrier was increased, which resulted in smaller gastric ulcer area in chalcone groups. these properties were also found to be dose dependent as they improved gastric mucosal state by increasing dosage. furthermore, this study revealed that administration of chalcone at low and high dose reduced ulcers in stomach. these findings were in line with the previous findings in terms of protective mechanisms of chalcone and flavonoids. the findings of this study suggest the use of chalcone with nsaids to reduce the risk of pud. also in a study by okunrobo and colleagues they tested the anti-inflammatory and gastro-protective properties of a synthetic chalcone with different dosages on rats who compared with the acetylsalicylic acid and cimetidine rat groups. the results were similar with the results present study terms of gastric wall mucosal characteristics and acidity of gastric content. finally omeprazole was used as antiulcer reference chemicals in this study against induced ulcer by indomethacin. omeprazole as a proton pump inhibitors are broadly prescribed medications with the control of gastric acid secretion for the prevention or treatment of peptic ulcer. omeprazole can suppress the gastric acid secretion through the h+,k + −atpase. it has been discovered that the h+,k + −atpase as gastric cell proton pump transfers h+ in exchange for luminal k+ to generate a highly acidic environment in the stomach that is responsible for gastric acid secretion. lastly indomethacin as a potent inhibitor of prostaglandins synthesis diminishes the protective effect of pge on mucosa of gastric wall.  acute toxicity tests did not show any sign of morbidity or mortality up to 1000 mg/kg of hptp. rats pre-treated with 50 mg/kg or 100 mg/kg of hptp before administration of 100 mg/kg of indomethacin significantly protect erosive gastric mucosal damage compared to ulcerogenic control group. histologic findings showed that hptp remarkably protect gastric mucosa compared to ulcerogenic control group. the gastroprotective effects of hptp could be attributed to increase in ph and mucus content of gastric content, increase endogenous enzymes and pge and decrease in mda level in gastric homogenate, over-expresion of hsp and tgf-β proteins, and down-expression of bax protein. 
this study has revealed that chalcone1--3-m-tolyl-propenone can serve as a safe and effective antiulcer agent as it has been proved to increase ph and gastric wall mucus, increase gpx, sod, pge and decrease mda level, ultimately, it has also contributes towards the over-expression of hsp protein andtgf-β, and down-expression of bax protein. pas staining showed intense uptake of magenta color of gastric wall mucus in rats fed with hptp, and immunohistochemical staining of gastric mucosa revealed over-expression of hsp protein, down-expression of bax protein and over expression of tgf-β in rats administered with hptp. administration of hptp had decreased the acidity, increased gastric wall mucus and flattening of gastric mucosa and reducing erosive gastric damage area. h & e stain showed decreased infiltration of leucocytes with edema of submucosal layer. non-steroidal anti-inflammatory drugs can result in peptic ulcer disease which is a common condition worldwide.
 non-steroidal anti-inflammatory drugs can result in peptic ulcer disease which is a common condition worldwide. the aim of this study was to evaluate the antiulcer properties of -3-m-tolyl-propenone) chalcone in rats using indomethacin as ulcerogenic agent.  none of the rats showed symptoms of kidney and liver toxicity during the term of the study. administration of hptp had decreased the acidity, increased gastric wall mucus and flattening of gastric mucosa and reducing erosive gastric damage area. hptp also showed dose dependent increase in sod, gpx activity and pge level and decrease mda. h & e stain showed decreased infiltration of leucocytes with edema of submucosal layer. pas staining showed intense uptake of magenta color of gastric wall mucus in rats fed with hptp, and immunohistochemical staining of gastric mucosa revealed over-expression of hsp protein, down-expression of bax protein and over expression of tgf-β in rats administered with hptp.  this study has revealed that chalcone1--3-m-tolyl-propenone can serve as a safe and effective antiulcer agent as it has been proved to increase ph and gastric wall mucus, increase gpx, sod, pge and decrease mda level, ultimately, it has also contributes towards the over-expression of hsp protein andtgf-β, and down-expression of bax protein. electronic supplementary material the online version of this article contains supplementary material, which is available to authorized users. keywords chalconeindomethacinpeptic ulcerantioxidantimmunohistochemistryhistologyendogenous enzymesissue-copyright-statement© the author 2014 