Roles of NADPH oxidase in occurrence of gastric damage and expression of cyclooxygenase-2 during ischemia/reperfusion in rat stomachs.

NADPH oxidase is an enzyme that converts molecular oxygen into reactive oxygen species, which cause severe damage in several organs. Cyclooxygenase (COX)-2 is an inducible enzyme that is important in gastric mucosal defense and repair processes. It is unclear whether NADPH oxidase is related to COX expression in the gastric mucosa, so we investigated the correlation. Under urethane anesthesia, a male Sprague Dawley rat stomach was mounted in an ex-vivo chamber, and ischemia/reperfusion (I/R) was performed through a cannula in the femoral vein. I/R significantly increased NADPH oxidase activity, H(2)O(2) production, and myeloperoxidase (MPO) activity. In contrast, ischemia alone clearly enhanced both NADPH oxidase activity and H(2)O(2) production but not MPO activity. Pretreatment with the NADPH oxidase inhibitor diphenylene iodonium (DPI) suppressed I/R-induced mucosal damage. On the other hand, the selective COX-2 inhibitor rofecoxib exhibited a tendency to enhance the severity of gastric damage induced by I/R, although the selective COX-1 inhibitor SC-560 and the nonselective COX inhibitor indomethacin had no effect. I/R also increased the expression of COX-2, and this increase was suppressed by pretreatment with DPI. These findings suggest that the increase in NADPH oxidase activity is involved in the occurrence of gastric mucosal damage induced by I/R and that this enzyme activity may be causally related to the upregulation of COX-2 during I/R.

Impairment by 5-fluorouracil of the healing of gastric lesions in rats: effect of lafutidine, a histamine H2 receptor antagonist, mediated by capsaicin-sensitive afferent neurons.

We investigated the influence of 5-fluorouracil (5-FU), an anti-tumor agent, on the healing of gastric lesions generated by 0.6 M HCl in rats and the effect of lafutidine, a histamine H(2) receptor antagonist, on the impaired healing. Animals fasted for 18 h were given 1 ml of 0.6 M HCl p.o., fed normally from 1 h later, and killed 1-96 h thereafter. 5-FU was given i.v. twice, 1 h and 24 h after the HCl. The gastric lesions healed spontaneously within 96 h. Although it decreased acid secretion, 5-FU markedly delayed the healing. Lafutidine, but not cimetidine, given p.o. immediately after each dosing of 5-FU significantly reversed the delay in healing by 5-FU, and this effect was attenuated by the chemical ablation of capsaicin-sensitive afferent neurons. Capsaicin also significantly reversed the delay in healing. The mucosal application of 50 mM HCl did not affect gastric mucosal blood flow (GMBF) in the normal stomach, but significantly increased it in the stomach damaged by 0.6 M HCl. The increases in GMBF were attenuated by 5-FU; however, the co-administration of lafutidine significantly restored the response. In addition, 5-FU inhibited both cell proliferation and migration in rat gastric epithelial cells (RGM1) in vitro. These results suggest that 5-FU delayed the healing of gastric lesions generated by 0.6 M HCl, probably through the inhibition of cell migration and proliferation, as well as the impairment of GMBF, and lafutidine reversed the delay in healing, mainly through the amelioration of the GMBF response mediated by capsaicin-sensitive afferent neurons.

Less irritative action of wine and Japanese sake in rat stomachs: a comparative study with ethanol.

The ingestion of alcohol, especially in excess, causes acute gastric lesions and gastritis in humans, yet the mucosal irritative action of alcoholic beverages remains largely unknown. We examined the mucosal irritative action of whiskey, wine and Japanese sake in the rat stomach both ex vivo and in vitro, in comparison with ethanol. Under urethane anesthesia, a rat stomach was mounted in an ex vivo chamber, then superfused with saline, and the transmucosal potential difference (PD) was measured. After the basal PD had stabilized, the mucosa was exposed for 30 min to 2 ml of 15% ethanol, whiskey (containing 15% ethanol), white wine, or Japanese sake (the ethanol concentration of the latter two is 12-15%). In the in vitro study, rat epithelial cells (RGM1) were treated with the alcoholic beverages for 5 min, and the cell viability was determined with crystal violet. Ethanol or whiskey applied to the chamber caused a decrease in PD, while wine or Japanese sake did not. Histologically, surface epithelial damage was observed after exposure to both ethanol and whiskey, yet no damage was induced by white wine and Japanese sake. Likewise, both ethanol and whiskey markedly reduced the viability of RGM1 cells after 5 min of incubation, while neither white wine nor Japanese sake had any effect. In addition, supplementation of glucose significantly prevented the reduction in both PD and cell viability caused by ethanol. These results suggest that the mucosal irritative action of Japanese sake and white wine is much less pronounced than that of ethanol or whiskey and that the less damaging action of Japanese sake and white wine may be, at least partly, accounted for by the glucose contained in these alcoholic beverages.

Irritative action of alcoholic beverages in rat stomachs: a comparative study with ethanol.

The mucosal irritative action of alcoholic beverages such as white wine, Japanese sake and whisky was examined in rat stomachs in vivo and in vitro, in comparison with ethanol. The concentration of ethanol in these alcoholic beverages was 15%. Mucosal application of ethanol (15%) and whisky in the chambered stomach caused a decrease in gastric potential difference (PD), while that of Japanese sake and white wine caused a slight increase but not decrease in PD. Likewise, both ethanol and whisky markedly reduced the cell viability of RGM1 cells after 5 min incubation, whereas neither Japanese sake nor white wine had any effect. In addition, supplementation of glucose, one of the non-alcoholic ingredients of white wine and Japanese sake, antagonized a reduction in both PD and cell viability caused by ethanol. These results suggest that the mucosal irritative action of Japanese sake and white wine is much less than that of ethanol or whisky and that these properties may be, at least partly, due to the glucose contained in these alcoholic beverages.

Prostaglandin E2 aggravates gastric mucosal injury induced by histamine in rats through EP1 receptors.

We demonstrated that prostaglandin (PG) E2 aggravates gastric mucosal injury caused by histamine in rats, and investigated using various EP agonists which EP receptor subtype is involved in this phenomenon. Rats were used after 18 hr fasting. Histamine (80 mg/kg) dissolved in 10% gelatin, was given s.c., either alone or in combination with i.v. administration of PGE2 or various EP agonists such as 17-phenyl PGE2 (EP1), butaprost (EP2), sulprostone (EP1/EP3), ONO-NT012 (EP3) and ONO-AE1-329 (EP4). The animals were killed 4 hr later, and the mucosa was examined for lesions. The mucosal permeability was determined using Evans blue (1%). Histamine alone induced few lesions in the gastric mucosa within 4 hr. PGE2 dose-dependently worsened the lesions induced by histamine, the response being inhibited by tripelennamine but not cimetidine. The effect of PGE2 was mimicked by 17-phenyl PGE2 and sulprostone, but not other EP agonists, including EP2, EP3, and EP3/EP4 agonists. The mucosal vascular permeability was slightly increased by histamine, and this response was markedly enhanced by co-administration of 17-phenyl PGE2 as well as PGE2. The mucosal ulcerogenic and vascular permeability responses induced by histamine plus PGE2 were both suppressed by pretreatment with ONO-AE829, the EP1 antagonist. These results suggest that PGE2 aggravates histamine-induced gastric mucosal injury in rats. This action of PGE2 is mediated by EP1 receptors and functionally associated with potentiation of the increased vascular permeability caused by histamine through stimulation of H1-receptors.

Prostaglandin E receptor EP1 subtype but not prostacyclin IP receptor involved in mucosal blood flow response of mouse stomachs following barrier disruption.

AIM: We investigated the role of prostacyclin IP receptors in gastric mucosal protection as well as functional responses induced by taurocholate Na (TC) as a mild irritant using IP receptor knockout mice, in comparison with prostaglandin (PG) E receptor EP1 subtype knockout animals. METHODS: Male C57/BL6 mice fasted for 18 h were used under urethane anesthesia. The stomach mounted on an ex vivo chamber was perfused with 20 mM HCl, and the transmucosal potential difference (PD), luminal acid loss, and gastric blood flow (GMBF) were simultaneously measured before and after exposure of the stomach to 20 mM TC for 20 min. RESULTS: Mucosal exposure to TC in wild-type mice caused a marked decrease in PD, followed by an increase in H(+) loss and GMBF. The PD gradually normalized after removal of TC from the chamber, with minimal damage in the mucosa 1 h later. The increase of GMBF following TC treatment was inhibited by indomethacin with no change in PD reduction or H(+) loss, resulting in severe lesions in the mucosa. None of these responses induced by TC were significantly altered in IP receptor knockout mice. However, in mice lacking EP1 receptors, TC did not increase GMBF, despite causing PD reduction and acid loss, and resulted in severe damage in the mucosa. Mucosal PGE(2) levels were significantly increased after TC, similarly, in all groups of mice, while levels of 6-keto-PGF(1alpha) showed a slight but not significant increase in all groups. CONCLUSION: We confirmed the importance of endogenous PGs and EP1 receptors in the adaptive protection and functional responses induced by a mild irritant in mouse stomach and further suggested that IP receptors are not actively involved in maintaining the gastric mucosal integrity under adverse conditions.