BACKGROUND
stomach ulcers are the common gastrointestinal and global disorders  <cit> .
it occurs mainly due to the imbalance between the destructive and offensive factors of the mucosal barrier  <cit> .
the destructive factors include stomach hydrochloric acid , mucosal hypoperfusion, free oxygen radicals, ethanol, helicobacter pylori and excessive ingestion of non-steroidal anti-inflammatory drugs  that promote the gastric mucosal injury and contribute to gastric ulceration  <cit> .
the prevention or treatment of gastric ulcers is a medical challenge  <cit> .
gastric ulcer therapy has major disadvantages, including limited efficacy of drugs against the gastric illness and severe side effects  <cit> .
therefore, medicinal plants may be a viable alternative therapy that has fewer side effects and contains a wide variety of antioxidants.
medicinal plants are also promising alternative medications for the development of new drugs to control gastrointestinal diseases, which has been reviewed extensively in the literature .
cibotium barometz hair  is known traditionally as “golden hair dog fern”.
cibotium is a tropical native medicinal plant in the malaysian peninsula and parts of china.
it is an anti-inflammatory plant and is used in contradiction of rheumatic and menstruation problems, as well as herniated discs and hyperosteogeny  <cit> .
there are many phenolic compounds in cibotium that are potent antioxidants and strong chelators.
the rhizomes are comprised of approximately β-sitosterol, caffeic acid, daucosterol, alternariol, 5-hydroxymethyl-2-furancarboxaldehyde, -des-o-methyl lasiodiplodin, protocatechuic aldehyde, -stigmast-4-ene-3-one, onitin, 24-methylenecycloartanol, protocatechuic acid, n-butyl-β-d fructopyranoside, palmitic acid, 1-monopalmitin, d-glucose and 30% starch  <cit> .
it has been confirmed that c. barometz obstructs osteoclast creation with no effects on cell viability  <cit> .
the hairof cibotium is a staple ingredient in the ointments that are applied in natural treatments, such as to stop bleeding  <cit> .
the objective of our study was to evaluate the stomach-protective effect of c. barometz hair ethanol extracts on ethanol-induced stomach ulcers in rats.
RESULTS
antiulcer study
gross estimations
the results that pre-treated with c. barometz hair ethanol extract of male sd rats shown significant reduced of the ulcer area with compared to the ulcer groups  at four different doses in the pre-treated animals groups  that were induced by ethanol 95%.
however, the significant inhibition percentage of the ulcer area increased in rats pre-treated with c. barometz hair at doses of  <dig>  mg\kg, 250 mg\kg, 125 mg\kg, and 500 mg\kg by  <dig> %,  <dig> %,  <dig> %, and  <dig> % respectively.table  <dig> effect of the c. barometz hair extracts on the mucus weight, ph of stomach content, ulcer area, and % inhibition of ulcer area in the stomach
*the values are expressed as the mean ± sem.
indicates significance at p <  <dig>  compared to the ulcerated group
fig.
<dig> the effect of c. barometz hair on the macroscopic appearance of the stomach mucosa in alcohol-induced stomach mucosal injuries in male sd rats.
g <dig>  exhibited no injuries to the gastric mucosa, g <dig>  had severe injuries to the stomach mucosa, g <dig>  showed mild disruptions of the surface epithelium in the gastric mucosa.
g <dig> , g <dig> , g <dig> , g <dig>  doses of c. barometz hair extract had moderate to mild disruptions of the surface epithelium in the gastric mucosa in a dose-dependent manner.
black arrow points to the hemorrhagic bands
gastric mucus content and acidity
the outcomes that shown in table  <dig>  the ulcerated sd rat group produced the lowest gastric mucosa mucus content, while animal groups pre-treated with g <dig>  and g <dig>  of c. barometz hair exhibited significant increasing in the mucus weight  with respect to g <dig> .
however, pre-treatment with c. barometz hair  produced a significant increase in the ph of the stomach contents compared to the ulcer control rats .
histological protocol of evaluation the stomach injuries
hematoxylin and eosin and periodic acid schiff  stainings
histology revealed comprehensive damage to the stomach mucosa in the ulcer control rats.
moreover, the ulcerated rat control group had necrotic lesions in the deep gastric mucosa that demonstrated extensive leucocyte infiltration and edema of the submucosal layer, as shown in fig.
<dig>  conversely, the animals pre-fed with c. barometz hair extract in the g <dig> - g <dig> groups presented relatively enhanced protection of the stomach mucosa with a depression in or lack of infiltration of leucocytes and edema .
c. barometz hair extracts revealed defending effects in a dose-dependent manner and showed remarkably improved protection of the stomach epithelium.
the gastric mucosa in the pre-treated experimental groups, depending on the dose, exhibited a gradual increase in pas staining intensity, indicated by the accumulation of the magenta color in the mucosal cell layer compared to the ulcer control group .
additionally, this magenta staining was reduced and was not plentiful in the gastric mucosa of the ulcer group where the ulcer was induced with ethanol.fig.
<dig> the effect of c. barometz on the histology  of ethanol-induced stomach mucosa damage in male sd rats.
g <dig>  had intact surface mucosal epithelium with no lesions; g <dig>  had a severe distraction of the surface epithelium and necrotic lesions; g <dig>  had a mild distraction of the surface epithelium and reduction in the submucosal edema with inflammatory cells infiltration.
the animals pretreated with c. barometz extract in g <dig> , g <dig> , g <dig>  and g <dig>  groups revealed a moderate to mild disruption of the surface epithelium, a reduction in submucosal edema and inflammatory cells infiltration in a dose-dependent manner as shown by lowering in or absence of the ulcer area , submucosal edema and inflammatory cells infiltration .
fig.
<dig> the effect of c. barometz on stomach tissue glycoprotein pas staining in ethanol-induced gastric ulcers in sd male rats.
g <dig>  had no accumulation of the magenta color in the mucosal cell layer; g <dig>  had decreased levels of the magenta color, g <dig> , g <dig> , g <dig> , g <dig> , and g <dig>  showed increases in pas staining intensity in mucosal cells layer compared to the ulcerated group, independent of dose.
the red arrow indicates the pas staining of glycoprotein.
immunohistochemistry
in the gastric mucosa, the expression of the hsp <dig> protein was down-regulated in the ulcerated group , but significantly up-regulated in the animals pre-treated with omeprazole  and with c. barometz hair extract , as displayed in fig.
<dig>  additionally, the immunohistochemical staining of the bax proteins  in the gastric mucosa revealed up-regulation in the ulcerated group while a significant down-regulation was demonstrated in rats pre-treated with c. barometz extract.fig.
<dig> immunohistochemistry study of hsp <dig> expression in the stomach mucosa of sd male rats.
g <dig> , g <dig> , g <dig> , g <dig> , g <dig> , g <dig>  and g <dig> .
hsp <dig> protein expression was up-regulated in animals pre-treated with c. barometz in a dose-dependent manner and hsp <dig> protein expression was also up-regulated with omeprazole.
down-regulation appeared in the ulcerated control group .
all data are stated as the mean ± sem.
the mean difference was significant at the p <  <dig>  level compared to the normal control group
fig.
<dig> immunohistochemistry study of bax expression the protein in the stomach mucosa of sd male rats.
g <dig> , g <dig> , g <dig> , g <dig> , g <dig> , g <dig>  and g <dig> .
bax protein expression was down-regulated in rats pre-treated with omeprazole and c. barometz in a dose-dependent manner.
bax was up-regulated in the ulcerated control group.
.
all data are stated as the mean ± sem.
the mean difference was significant at the p <  <dig>  level compared to the normal control group
measurement of antioxidant enzymes and membrane lipid peroxidation  of stomach
a significant reduction was observed in endogenous antioxidants enzymes  activities for the ulcer group of male sd rats.
however, the rats pre-treated with c. barometz hair displayed an elevation of all antioxidant activities with respect to the  ulcer group as shown in figs.
6a, b, and c. the sod enzyme activities were significantly higher at doses for g <dig> of c. barometz than g <dig> .
the cat enzyme activities illustrate the significant increases in g <dig>  g <dig>  g <dig> groups compared to g <dig> as shown in fig.
6b.
the gpx enzyme activities for gastric mucosal homogenates revealed significant increasing in the rats pre-fed with four doses of c. barometz hair ethanol extract with respect to g <dig> as shown in fig.
6c.
additionally, the mda level of c. barometz hair extract in g <dig>  g <dig>  g <dig> and g <dig> were significant lower than in the g <dig> ulcerated control group as seen in fig.
6d.fig.
<dig> effect of c. barometz hair ethanol extracts on gastric tissue homogenate of superoxide dismutase , catalase , glutathione  and malondialdehyde  assays.
g <dig> , g <dig> , g <dig> , g <dig> , g <dig> , g <dig>  and g <dig>  of c. barometz hair ethanol extract.
all values  are expressed as the mean ± sem.
* significant at p < <dig>  compared to ulcerated group.
sod, cat and gpx, antioxidant activities were higher in g <dig>  g <dig> and g <dig> to g <dig> than in g <dig>  although the mda levels of g <dig> were higher than those of the other groups
acute toxicity test of c. barometz hair
all sd rats that were treated with c. barometz hair ethanol extract demonstrated no mortality and toxic signs in the experiment.
there were no signs of hepatotoxic or nephrotoxic effects, which were evaluated histologically and biochemically .
additionally, there were no body weight variations or abnormal physiological or behavioral changes at 2 g/kg and 5 g/kg doses during the 14 days, compared to the control group that was given 10% tween  <dig> fig.
<dig> histological slides of the liver and kidney sections for the acute toxicity experiment.
animals treated with 5 ml/kg of the vehicle  a and b. animals treated with 2000 mg/kg  of the c. barometz hair extract c and d. animals treated with 5000 mg/kg  of the c. barometz hair extract e and f. no significant changes were observed in the structures of the liver and kidney among the treated and control groups
antioxidants activities of ethanol extract of c. barometz hair
ferric reducing antioxidant power  test
c. barometz hair antioxidant activity in vitro was measured totally using the frap test.
fig.
<dig> revealed the decline of ferric to ferrous ions that indicated a higher frap value for c. barometz hair /g) than bht /g) and ascorbic acid /g).
however, the value was less than for quercetin /g) and gallic acid /g) standards.fig.
<dig> frap test for total antioxidant activity evaluation of c. barometz hair extract with reference synthetic standards  were determined.
all values are in triplicate and represented as the mean ± sem
the scavenging of diphenyl - picrylhydrazyl radical activity  test
c. barometz hair scavenging of dpph free radicals was assessed using the dpph test.
fig.
<dig> demonstrated the inhibition% of the dpph free-radical scavenging activity of c. barometz hair that was  <dig> % with an ic <dig> value of  <dig>  ±  <dig>  μg/ml.
it was associated to the standards bht, ascorbic acid, quercetin and gallic acid.
the % inhibition of dpph free-radicals scavenging activities of the standards was  <dig> ,  <dig> ,  <dig> , and  <dig> % with an ic <dig> value of  <dig>  ±  <dig>  μg/ml,  <dig>  ±  <dig>  μg/ml,  <dig>  ±  <dig>  μg/ml,  <dig>  ±  <dig>  μg/ml, respectively.fig.
<dig> dpph test for free radical scavenging activity  of c. barometz hair ethanol extract with reference synthetic standards .
all values are in triplicate and represented as the mean ± sem
discussion
traditional medicine has become an important type of alternative medicine in many countries.
several medicinal plants have been used to protect the stomach from a number of ulcerative agents.
one type of medicinal plant is c. barometz leaves, which are used to stop bleeding  <cit> .
in this study, the results of acute toxicity experiment revealed no signs of toxicity or mortality in vivo and no variations in kidney and liver on biochemical investigation.
additionally, no differences in body weights were observed for both sex sd rats.
these findings corroborate previous studies .
our data suggest that c. barometz hair extract has high free-radicals scavengings and antioxidant activities in vitro.
our results agree with a previous report that indicated that the consumption of medicinal plants that contain natural antioxidants can reduce free radicals and protect biological molecules from oxidative injury  <cit> .
c. barometz hair is one of the medicinal plants that have medicinal properties and natural antioxidants  <cit> .
c. barometz hair displays good free-radical scavenging activity and ferric-decreasing antioxidant power in a fe3+-dependent hydroxyl-radical generation assay  <cit> .
this outcome agrees with earlier studies of c. barometz, in which it showed a greater antioxidant activity  <cit> .
our data indicate that significant protection of stomach wall mucosa and decreases in ulcer area are observed in rats pre-fed with c. barometz extract.
our observation is consistent with the findings of several other studies  <cit> .
we also observed a significant elevation of stomach mucus content and ph in rats pre-fed with c. barometz hair extract compared to category of the ulcer control rats.
this shows the extracted plant of c. barometz is able to protect the mucosal layer of the stomach from destructive causes.
these results are similar to those previously reported by others .
a severe distraction of the gastric mucosa layer, edema and leucocyte penetration of submucosal layers were apparent in the ulcer control rats compared with rats pre-fed with c. barometz hair extract.
the rats treated with c. barometz hair extract displayed a mild distraction of stomach epithelium and reduced edema and inflammatory cell penetration of the submucosal layers.
similarly, many studies have also observed mild distraction of stomach mucosa, mild edema and penetration of inflammatory cells in the submucosal layers .
additionally, the condensing of the submucosal layer was detected in the ulcerated group where there was edema and hemorrhagic abrasions in the mucosal layer, which is an indicator of ethanol damage.
absolute ethanol harshly damages the stomach glandular epithelium leading to augmented neutrophils access into the disrupted stomach mucosa.
the presence of oxygen free radicals resulting from the penetration of neutrophils in the wounded gastric wall can damage stomach gastric mucosa in rats  <cit> .
neutrophils are the main sources of inflammatory mediators and can release the potent reactive oxygen species, which are cytotoxic and can promote tissue injury  <cit> .
furthermore, neutrophils that congregate in the stomach mucosa have been exposed to irritation through the microcirculatory defect  <cit> .
the present study illustrated strong staining of glycoprotein discharge of stomach glandular epithelium in rats pre-fed with c. barometz extract.
mucus released from the stomach epithelium performs one of the vital mechanism of stomach mucosal barrier resistance against absolute ethanol  <cit> .
mucus and bicarbonate liberation may have a considerable role in ulcer development because the mucus/bicarbonate layers guards newly produced cells from acidic and peptic damage  <cit> .
the amplified production of mucus in the present work was an indicator of local mucosal defense in the stomach and can be described as the possible cytoprotective mechanism.
numerous studies have suggested that the gastroprotective defense is due to increased mucosal resistance, and a decline in aggressive factors, primarily acid and pepsin .
the consequences of the present immunohistochemistry experiments indicate that rats pre-fed with c. barometz hair displayed an up-regulation of hsp <dig> proteins that contributed to the protection of stomach cells from heat shock, oxidative stress, and down-regulation of the bax proteins.
earlier studies reported that there was up-regulation of hsp <dig> protein in animals fed with plant extracts  <cit> .
however, the down-regulation of heat shock protein expression appeared in the ulcer control animals as a gastric injury marker.
this is consistent with the results from other studies  <cit> .
up-regulation of hsp <dig> protein in the stomach wall mucosa in animals pre-fed with c. barometz hair extract may stop the initiation of damage to the gastric epithelium upon exposure to absolute alcohol  <cit> .
the hsp <dig> protein protected mitochondria and delayed a stress-induced apoptotic programmer  <cit> .
our data suggest that bax immune-staining confirmed that rats fed with c. barometz hair extract displayed increased an expression of the bax protein.
thus, these results demonstrate that c. barometz hair extract is a protective agent and prevents alcohol-induced damage to the rat stomach, which is associated with decreased expression of the bax protein.
similarly, many amendments have been described by many investigators  <cit> .
animals pre-fed with c. barometz hair extract displayed decreased expression of the bax protein.
on the other hand, up-regulation of bax was observed in the ulcer control group.
bax is a key protein that is linked to apoptosis during mitochondrial damage and plays a significant role in the disruption of stomach mucosal integrity that is observed after ethanol administration  <cit> .
the results from the present work demonstrated that rats pre-fed with c. barometz displayed an increase in the activity of endogenous antioxidant enzymes  and a decrease in mda levels in the stomach compared to the ulcer control rats.
these findings are consistent with observations from other studies  <cit> .
therefore, the elevation of sod and gpx enzymatic activities leads to increased scavenging of superoxides, hydrogen peroxide, hydroxyl and lipid peroxyl radicals, resulting in a reduction in tissue damage  <cit> .
additionally, increases in cat enzymatic antioxidant activity rapidly converted the peroxyl radicals into biologically safe substance, such as water  <cit> .
in contrast, the low levels of mda reduced lipid peroxidation and ros  which are products of oxidative gastric damage  <cit> .
taken together, our observations suggest that c. barometz hair facilitates gastric mucosa protection due to its scavenging of free radicals and anti-inflammatory effects  <cit> .
CONCLUSIONS
our study demonstrates that c. barometz hair has beneficial and dose-dependent anti-ulcer effects against ethanol-induced acute stomach hemorrhagic injuries in sd rats.
the gastro-protective effects of this effective medicinal plant could be associated with free radicals scavenging activities, elevated levels of ph and gastric mucus glycoprotein, increases in the cellular endogenous enzymes activity of sod, cat and gpx antioxidants, and a reduction in mda levels.
additionally, these effects may be due to hsp <dig> up-regulation and bax protein down-regulation.
thus, c. barometz hair is a promising gastro-protective agent that can potentially be used to treat gastric ulcers.

 stomach ulcers are the common gastrointestinal and global disorders. it occurs mainly due to the imbalance between the destructive and offensive factors of the mucosal barrier. the destructive factors include stomach hydrochloric acid, mucosal hypoperfusion, free oxygen radicals, ethanol, helicobacter pylori and excessive ingestion of non-steroidal anti-inflammatory drugs that promote the gastric mucosal injury and contribute to gastric ulceration. the prevention or treatment of gastric ulcers is a medical challenge. gastric ulcer therapy has major disadvantages, including limited efficacy of drugs against the gastric illness and severe side effects. therefore, medicinal plants may be a viable alternative therapy that has fewer side effects and contains a wide variety of antioxidants. medicinal plants are also promising alternative medications for the development of new drugs to control gastrointestinal diseases, which has been reviewed extensively in the literature. cibotium barometz hair is known traditionally as “golden hair dog fern”. cibotium is a tropical native medicinal plant in the malaysian peninsula and parts of china. it is an anti-inflammatory plant and is used in contradiction of rheumatic and menstruation problems, as well as herniated discs and hyperosteogeny. there are many phenolic compounds in cibotium that are potent antioxidants and strong chelators. the rhizomes are comprised of approximately β-sitosterol, caffeic acid, daucosterol, alternariol, 5-hydroxymethyl-2-furancarboxaldehyde, -des-o-methyl lasiodiplodin, protocatechuic aldehyde, -stigmast-4-ene-3-one, onitin, 24-methylenecycloartanol, protocatechuic acid, n-butyl-β-d fructopyranoside, palmitic acid, 1-monopalmitin, d-glucose and 30% starch. it has been confirmed that c. barometz obstructs osteoclast creation with no effects on cell viability. the hairof cibotium is a staple ingredient in the ointments that are applied in natural treatments, such as to stop bleeding. the objective of our study was to evaluate the stomach-protective effect of c. barometz hair ethanol extracts on ethanol-induced stomach ulcers in rats.  antiulcer study gross estimations the results that pre-treated with c. barometz hair ethanol extract of male sd rats shown significant reduced of the ulcer area with compared to the ulcer groups at four different doses in the pre-treated animals groups that were induced by ethanol 95%. however, the significant inhibition percentage of the ulcer area increased in rats pre-treated with c. barometz hair at doses of  mg\kg, 250 mg\kg, 125 mg\kg, and 500 mg\kg by %, %, %, and % respectively.table effect of the c. barometz hair extracts on the mucus weight, ph of stomach content, ulcer area, and % inhibition of ulcer area in the stomach *the values are expressed as the mean ± sem. indicates significance at p <  compared to the ulcerated group fig. the effect of c. barometz hair on the macroscopic appearance of the stomach mucosa in alcohol-induced stomach mucosal injuries in male sd rats. g exhibited no injuries to the gastric mucosa, g had severe injuries to the stomach mucosa, g showed mild disruptions of the surface epithelium in the gastric mucosa. g, g, g, g doses of c. barometz hair extract had moderate to mild disruptions of the surface epithelium in the gastric mucosa in a dose-dependent manner. black arrow points to the hemorrhagic bands gastric mucus content and acidity the outcomes that shown in table  the ulcerated sd rat group produced the lowest gastric mucosa mucus content, while animal groups pre-treated with g and g of c. barometz hair exhibited significant increasing in the mucus weight with respect to g. however, pre-treatment with c. barometz hair produced a significant increase in the ph of the stomach contents compared to the ulcer control rats. histological protocol of evaluation the stomach injuries hematoxylin and eosin and periodic acid schiff stainings histology revealed comprehensive damage to the stomach mucosa in the ulcer control rats. moreover, the ulcerated rat control group had necrotic lesions in the deep gastric mucosa that demonstrated extensive leucocyte infiltration and edema of the submucosal layer, as shown in fig.  conversely, the animals pre-fed with c. barometz hair extract in the g - g groups presented relatively enhanced protection of the stomach mucosa with a depression in or lack of infiltration of leucocytes and edema. c. barometz hair extracts revealed defending effects in a dose-dependent manner and showed remarkably improved protection of the stomach epithelium. the gastric mucosa in the pre-treated experimental groups, depending on the dose, exhibited a gradual increase in pas staining intensity, indicated by the accumulation of the magenta color in the mucosal cell layer compared to the ulcer control group. additionally, this magenta staining was reduced and was not plentiful in the gastric mucosa of the ulcer group where the ulcer was induced with ethanol.fig. the effect of c. barometz on the histology of ethanol-induced stomach mucosa damage in male sd rats. g had intact surface mucosal epithelium with no lesions; g had a severe distraction of the surface epithelium and necrotic lesions; g had a mild distraction of the surface epithelium and reduction in the submucosal edema with inflammatory cells infiltration. the animals pretreated with c. barometz extract in g, g, g and g groups revealed a moderate to mild disruption of the surface epithelium, a reduction in submucosal edema and inflammatory cells infiltration in a dose-dependent manner as shown by lowering in or absence of the ulcer area, submucosal edema and inflammatory cells infiltration. fig. the effect of c. barometz on stomach tissue glycoprotein pas staining in ethanol-induced gastric ulcers in sd male rats. g had no accumulation of the magenta color in the mucosal cell layer; g had decreased levels of the magenta color, g, g, g, g, and g showed increases in pas staining intensity in mucosal cells layer compared to the ulcerated group, independent of dose. the red arrow indicates the pas staining of glycoprotein. immunohistochemistry in the gastric mucosa, the expression of the hsp protein was down-regulated in the ulcerated group, but significantly up-regulated in the animals pre-treated with omeprazole and with c. barometz hair extract, as displayed in fig.  additionally, the immunohistochemical staining of the bax proteins in the gastric mucosa revealed up-regulation in the ulcerated group while a significant down-regulation was demonstrated in rats pre-treated with c. barometz extract.fig. immunohistochemistry study of hsp expression in the stomach mucosa of sd male rats. g, g, g, g, g, g and g. hsp protein expression was up-regulated in animals pre-treated with c. barometz in a dose-dependent manner and hsp protein expression was also up-regulated with omeprazole. down-regulation appeared in the ulcerated control group. all data are stated as the mean ± sem. the mean difference was significant at the p <  level compared to the normal control group fig. immunohistochemistry study of bax expression the protein in the stomach mucosa of sd male rats. g, g, g, g, g, g and g. bax protein expression was down-regulated in rats pre-treated with omeprazole and c. barometz in a dose-dependent manner. bax was up-regulated in the ulcerated control group.. all data are stated as the mean ± sem. the mean difference was significant at the p <  level compared to the normal control group measurement of antioxidant enzymes and membrane lipid peroxidation of stomach a significant reduction was observed in endogenous antioxidants enzymes activities for the ulcer group of male sd rats. however, the rats pre-treated with c. barometz hair displayed an elevation of all antioxidant activities with respect to the ulcer group as shown in figs. 6a, b, and c. the sod enzyme activities were significantly higher at doses for g of c. barometz than g. the cat enzyme activities illustrate the significant increases in g g g groups compared to g as shown in fig. 6b. the gpx enzyme activities for gastric mucosal homogenates revealed significant increasing in the rats pre-fed with four doses of c. barometz hair ethanol extract with respect to g as shown in fig. 6c. additionally, the mda level of c. barometz hair extract in g g g and g were significant lower than in the g ulcerated control group as seen in fig. 6d.fig. effect of c. barometz hair ethanol extracts on gastric tissue homogenate of superoxide dismutase, catalase, glutathione and malondialdehyde assays. g, g, g, g, g, g and g of c. barometz hair ethanol extract. all values are expressed as the mean ± sem. * significant at p < compared to ulcerated group. sod, cat and gpx, antioxidant activities were higher in g g and g to g than in g although the mda levels of g were higher than those of the other groups acute toxicity test of c. barometz hair all sd rats that were treated with c. barometz hair ethanol extract demonstrated no mortality and toxic signs in the experiment. there were no signs of hepatotoxic or nephrotoxic effects, which were evaluated histologically and biochemically. additionally, there were no body weight variations or abnormal physiological or behavioral changes at 2 g/kg and 5 g/kg doses during the 14 days, compared to the control group that was given 10% tween fig. histological slides of the liver and kidney sections for the acute toxicity experiment. animals treated with 5 ml/kg of the vehicle a and b. animals treated with 2000 mg/kg of the c. barometz hair extract c and d. animals treated with 5000 mg/kg of the c. barometz hair extract e and f. no significant changes were observed in the structures of the liver and kidney among the treated and control groups antioxidants activities of ethanol extract of c. barometz hair ferric reducing antioxidant power test c. barometz hair antioxidant activity in vitro was measured totally using the frap test. fig.  revealed the decline of ferric to ferrous ions that indicated a higher frap value for c. barometz hair /g) than bht /g) and ascorbic acid /g). however, the value was less than for quercetin /g) and gallic acid /g) standards.fig. frap test for total antioxidant activity evaluation of c. barometz hair extract with reference synthetic standards were determined. all values are in triplicate and represented as the mean ± sem the scavenging of diphenyl - picrylhydrazyl radical activity test c. barometz hair scavenging of dpph free radicals was assessed using the dpph test. fig.  demonstrated the inhibition% of the dpph free-radical scavenging activity of c. barometz hair that was % with an ic value of  ±   μg/ml. it was associated to the standards bht, ascorbic acid, quercetin and gallic acid. the % inhibition of dpph free-radicals scavenging activities of the standards was,,, and % with an ic value of  ±   μg/ml,  ±   μg/ml,  ±   μg/ml,  ±   μg/ml, respectively.fig. dpph test for free radical scavenging activity of c. barometz hair ethanol extract with reference synthetic standards. all values are in triplicate and represented as the mean ± sem discussion traditional medicine has become an important type of alternative medicine in many countries. several medicinal plants have been used to protect the stomach from a number of ulcerative agents. one type of medicinal plant is c. barometz leaves, which are used to stop bleeding. in this study, the results of acute toxicity experiment revealed no signs of toxicity or mortality in vivo and no variations in kidney and liver on biochemical investigation. additionally, no differences in body weights were observed for both sex sd rats. these findings corroborate previous studies. our data suggest that c. barometz hair extract has high free-radicals scavengings and antioxidant activities in vitro. our results agree with a previous report that indicated that the consumption of medicinal plants that contain natural antioxidants can reduce free radicals and protect biological molecules from oxidative injury. c. barometz hair is one of the medicinal plants that have medicinal properties and natural antioxidants. c. barometz hair displays good free-radical scavenging activity and ferric-decreasing antioxidant power in a fe3+-dependent hydroxyl-radical generation assay. this outcome agrees with earlier studies of c. barometz, in which it showed a greater antioxidant activity. our data indicate that significant protection of stomach wall mucosa and decreases in ulcer area are observed in rats pre-fed with c. barometz extract. our observation is consistent with the findings of several other studies. we also observed a significant elevation of stomach mucus content and ph in rats pre-fed with c. barometz hair extract compared to category of the ulcer control rats. this shows the extracted plant of c. barometz is able to protect the mucosal layer of the stomach from destructive causes. these results are similar to those previously reported by others. a severe distraction of the gastric mucosa layer, edema and leucocyte penetration of submucosal layers were apparent in the ulcer control rats compared with rats pre-fed with c. barometz hair extract. the rats treated with c. barometz hair extract displayed a mild distraction of stomach epithelium and reduced edema and inflammatory cell penetration of the submucosal layers. similarly, many studies have also observed mild distraction of stomach mucosa, mild edema and penetration of inflammatory cells in the submucosal layers. additionally, the condensing of the submucosal layer was detected in the ulcerated group where there was edema and hemorrhagic abrasions in the mucosal layer, which is an indicator of ethanol damage. absolute ethanol harshly damages the stomach glandular epithelium leading to augmented neutrophils access into the disrupted stomach mucosa. the presence of oxygen free radicals resulting from the penetration of neutrophils in the wounded gastric wall can damage stomach gastric mucosa in rats. neutrophils are the main sources of inflammatory mediators and can release the potent reactive oxygen species, which are cytotoxic and can promote tissue injury. furthermore, neutrophils that congregate in the stomach mucosa have been exposed to irritation through the microcirculatory defect. the present study illustrated strong staining of glycoprotein discharge of stomach glandular epithelium in rats pre-fed with c. barometz extract. mucus released from the stomach epithelium performs one of the vital mechanism of stomach mucosal barrier resistance against absolute ethanol. mucus and bicarbonate liberation may have a considerable role in ulcer development because the mucus/bicarbonate layers guards newly produced cells from acidic and peptic damage. the amplified production of mucus in the present work was an indicator of local mucosal defense in the stomach and can be described as the possible cytoprotective mechanism. numerous studies have suggested that the gastroprotective defense is due to increased mucosal resistance, and a decline in aggressive factors, primarily acid and pepsin. the consequences of the present immunohistochemistry experiments indicate that rats pre-fed with c. barometz hair displayed an up-regulation of hsp proteins that contributed to the protection of stomach cells from heat shock, oxidative stress, and down-regulation of the bax proteins. earlier studies reported that there was up-regulation of hsp protein in animals fed with plant extracts. however, the down-regulation of heat shock protein expression appeared in the ulcer control animals as a gastric injury marker. this is consistent with the results from other studies. up-regulation of hsp protein in the stomach wall mucosa in animals pre-fed with c. barometz hair extract may stop the initiation of damage to the gastric epithelium upon exposure to absolute alcohol. the hsp protein protected mitochondria and delayed a stress-induced apoptotic programmer. our data suggest that bax immune-staining confirmed that rats fed with c. barometz hair extract displayed increased an expression of the bax protein. thus, these results demonstrate that c. barometz hair extract is a protective agent and prevents alcohol-induced damage to the rat stomach, which is associated with decreased expression of the bax protein. similarly, many amendments have been described by many investigators. animals pre-fed with c. barometz hair extract displayed decreased expression of the bax protein. on the other hand, up-regulation of bax was observed in the ulcer control group. bax is a key protein that is linked to apoptosis during mitochondrial damage and plays a significant role in the disruption of stomach mucosal integrity that is observed after ethanol administration. the results from the present work demonstrated that rats pre-fed with c. barometz displayed an increase in the activity of endogenous antioxidant enzymes and a decrease in mda levels in the stomach compared to the ulcer control rats. these findings are consistent with observations from other studies. therefore, the elevation of sod and gpx enzymatic activities leads to increased scavenging of superoxides, hydrogen peroxide, hydroxyl and lipid peroxyl radicals, resulting in a reduction in tissue damage. additionally, increases in cat enzymatic antioxidant activity rapidly converted the peroxyl radicals into biologically safe substance, such as water. in contrast, the low levels of mda reduced lipid peroxidation and ros which are products of oxidative gastric damage. taken together, our observations suggest that c. barometz hair facilitates gastric mucosa protection due to its scavenging of free radicals and anti-inflammatory effects.  our study demonstrates that c. barometz hair has beneficial and dose-dependent anti-ulcer effects against ethanol-induced acute stomach hemorrhagic injuries in sd rats. the gastro-protective effects of this effective medicinal plant could be associated with free radicals scavenging activities, elevated levels of ph and gastric mucus glycoprotein, increases in the cellular endogenous enzymes activity of sod, cat and gpx antioxidants, and a reduction in mda levels. additionally, these effects may be due to hsp up-regulation and bax protein down-regulation. thus, c. barometz hair is a promising gastro-protective agent that can potentially be used to treat gastric ulcers. 
homogenates of the stomach tissue demonstrated significant increases in the endogenous antioxidant enzymatic activity and decreased lipid peroxidation in rats pre-treated with c. barometz hair extract compared with the ulcer control rats. 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 hsp and down-regulation of bax proteins. 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. immunohistochemical analysis revealed that rats pre-fed with the plant extract showed an up-regulation of the heat shock protein and down-regulation of bax proteins compared to ulcer control rats. 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 rats were administered 10% tween in the normal control and ulcer control groups, and omeprazole 20 mg/kg and, and 500 mg/kg of c. barometz hair extract in the experimental groups. a periodic acid schiff 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. rats 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. after 60 min, the normal control group of rats was orally administered 10% tween while absolute ethanol was orally administered to the groups of ulcer control, omeprazole and experimental groups. cibotium barometz is a medical herb used traditionally in the malaysian peninsula for several ailments, including gastric ulcer.
 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 rats were administered 10% tween in the normal control and ulcer control groups, and omeprazole 20 mg/kg and, 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 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.  rats 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 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 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 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.  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 hsp and down-regulation of bax proteins. issue-copyright-statement© the author 2017 