Departments of 1 Applied Pharmacology and 2 Biopharmaceutics, Kyoto Pharmaceutical University, Kyoto 607-8414, Japan
![]() |
ABSTRACT |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Cyclooxygenase (COX)-2
expression is induced in the gastric mucosa of Helicobacter
pylori-infected patients, but its role remains unclear. We
examined the effects of NS-398 and indomethacin on gastric pathology in
H. pylori-infected Mongolian gerbils. COX-1 was detected in
both normal and H. pylori-infected mucosa, whereas COX-2 was
expressed only in the infected mucosa. PGE2 production was
elevated by H. pylori infection, and the increased production was reduced by NS-398, which did not affect PGE2
production in normal mucosa. Indomethacin inhibited PGE2
production in both normal and infected mucosa. Hemorrhagic erosions,
neutrophil infiltration, lymphoid follicles, and epithelium damage were
induced by H. pylori infection. NS-398 and indomethacin
aggravated these pathological changes but did not increase viable
H. pylori number. H. pylori-increased production
of neutrophil chemokine and interferon- was potentiated by NS-398
and indomethacin. Neither NS-398 nor indomethacin caused any
pathological changes or cytokine production in normal animals. These
results indicate that COX-2 as well as COX-1 might play anti-inflammatory roles in H. pylori-induced gastritis.
prostaglandin; nonsteroidal anti-inflammatory drug; infection
![]() |
INTRODUCTION |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
HELICOBACTER PYLORI is recognized as a major etiologic factor of chronic gastritis and gastric/duodenal ulcers. H. pylori infection causes marked infiltration of inflammatory cells into the gastric mucosa and expression of many cytokines. Consequently, it is generally suspected that cytokine-related inflammatory responses might be involved in H. pylori-induced mucosal injuries (3, 5, 10, 43). Similarly, mucosal defense mechanisms might also be activated by H. pylori infection. Prostaglandins (PGs) are well-known mucosal defense factors, protecting the gastric mucosa against injury caused by a variety of toxic stimuli (9, 28). PGE2 stimulates the secretion of gastric mucus and bicarbonate, increases mucosal blood flow, inhibits acid secretion, and reduces gastric motility (9). PGs are synthesized through cyclooxygenase (COX), which is a target of nonsteroidal anti-inflammatory drugs (NSAIDs) (38). COX exists in two isoforms, of which COX-1 is constitutively expressed in many tissues, including the stomach, and COX-2 is normally undetectable in most tissues, its expression being induced at inflammatory sites (13, 23). In the normal gastric mucosa, COX-1-derived PGs play a crucial role in maintaining mucosal integrity (14, 18). In contrast, COX-2 is not expressed in normal mucosa, but its induction results in a marked and sustained increase in PGE2 production in the damaged mucosa (24, 32, 37). In addition to COX-1, COX-2 also plays an important role in the healing of gastric ulcers (24, 32). Recent studies showed that COX-2 expression is observed in the gastric mucosa of H. pylori-positive patients (22, 31, 33). In addition, Romano et al. (29) reported that H. pylori directly induces COX-2 mRNA expression and increases PGE2 production in gastric cancer MKN28 cells. However, the role of COX-2 in H. pylori-induced gastritis in in vivo remains unclear.
Mongolian gerbil models of H. pylori infection have been established and widely used (15, 16, 20, 21, 34). We have shown that chronic gastritis and ulcers are generated in all H. pylori-infected animals (20, 34). In addition, Watanabe et al. (41) recently reported that gastric cancers are also developed by H. pylori infection in gerbils. Thus the gerbil model is quite suitable for in vivo study of the pathogenesis of H. pylori-induced gastric diseases, since this model exhibits pathological features that mimic those of human patients with H. pylori.
To investigate the role of COX-2 in H. pylori-induced gastritis, we examined the effect of NS-398, a COX-2-selective inhibitor, on H. pylori-induced pathological changes in Mongolian gerbils, compared with indomethacin, a nonselective COX inhibitor.
![]() |
METHODS |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Animals. Male Mongolian gerbils (6 wk old, 40-50 g) were kindly supplied from Nihon SLC (Hamamatsu, Japan). The animals were kept in an isolated clean room with regulated temperature (20-22°C) and humidity (~55%) with a 12:12-h light/dark cycle. The animals were fasted for 24 h before H. pylori inoculation, and drinking water was also withheld after the inoculation. From 4 h after the inoculation, both food and water were freely available to the animals.
H. pylori preparation and inoculation of Mongolian gerbils. The preparation and inoculation of H. pylori were performed as we previously described (34). A cagA- and vacA-positive standard strain of H. pylori (NCTC 11637; American Type Culture Collection, Rockville, MD) was used. The bacteria were incubated in a brain-heart infusion broth (Difco Laboratories, Detroit, MI) containing 10% fetal bovine serum (GIBCO BRL, Gaithersburg, MD) at 37°C overnight under a microaerophilic atmosphere and allowed to grow to a density of ~2.0 × 108 colony-forming units (CFU)/ml. H. pylori (2.0 × 108 CFU; 1.0 ml) was orally administered to each animal. Normal animals received 1.0 ml of the medium alone.
Western blot analysis of COX-1 and COX-2 proteins. Expression of COX-1 and COX-2 proteins in the gastric mucosa was examined by Western blotting. Gastric specimens (the fundus near the antrum) were taken from normal and H. pylori-infected animals. COX proteins were partially purified according to the method of Gierse et al. (12). The specimens were homogenized in 25 mM Tris · HCl (pH 8.0) buffer containing 250 mM sucrose, followed by centrifugation at 10,000 g for 20 min. The pellet was resuspended in 25 mM Tris · HCl (pH 8.0) buffer containing 1% 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, and the mixture was gently stirred for 2 h at 4°C. The supernatant was recovered after centrifugation at 30,000 g for 30 min and applied onto a DEAE-Sepharose CL-4B (Amersham Pharmacia Biotech, Little Chalfont, UK) column that had been equilibrated with 25 mM Tris · HCl (pH 8.0) buffer containing 0.5% 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, 1 mM phenylmethylsulfonyl fluoride, and 0.2 mM EDTA. After the column was washed with the same buffer supplemented with 50 mM NaCl, elution was performed with 200 mM NaCl. After aliquots (20 µg) of the eluted proteins and purified COX proteins (50 ng; Cayman, Ann Arbor, MI) had been subjected to SDS-polyacrylamide gel electrophoresis (10%), the separated proteins were electrophoretically transferred onto Hybond-P membranes (Amersham Pharmacia Biotech). The membranes were incubated with the antibody (Cayman) against COX-1 or COX-2 protein after nonspecific binding sites had been blocked with bovine serum albumin. COX proteins were detected on X-ray films (Fuji Film, Tokyo, Japan) with an enhanced chemiluminescence kit (Immuno Star; Wako Pure Chemicals, Osaka, Japan).
Determination of PGE2 production. Gastric specimens (the fundus near the antrum) were taken from normal and H. pylori-infected animals. After being washed with PBS, the tissues were minced and then incubated in 1 ml of Dulbecco's modified Eagle's medium supplemented with 2.5% fetal bovine serum at 37°C for 1 h under 5% CO2 in air. The amount of PGE2 in the medium was determined by enzyme immunoassay (PGE2 EIA kit; Cayman). PGE2 production was expressed as picograms of PGE2 per milligram of tissue per hour.
Evaluation of H. pylori-induced gastritis. Gastric pathology was blindly evaluated. Normal and H. pylori-infected animals were killed, and their stomachs were excised. The stomachs were incised along the greater curvature and spread out with pins on a cork board. Gastric mucosal hemorrhagic lesions were examined under a dissecting microscope (magnification, ×10). Thereafter, four gastric specimens were cut off from the fundus near the antrum and fixed in 4% paraformaldehyde in PBS. Frozen sections (12 µm thick) were prepared, and neutrophil-specific myeloperoxidase activity-dependent staining was carried out (11). In brief, the sections were incubated in 50 mM Tris · HCl (pH 7.6) containing 0.2 mg/ml 3',3'-diaminobenzidine tetrahydrochloride (Dojindo, Kumamoto, Japan) in the presence of 0.005% H2O2 at room temperature. After being washed with PBS, the sections were successively stained with hematoxylin. Brown- or black-stained cells in the mucosa were identified as neutrophils when their morphological features were confirmed under a light microscope at high magnification and the cells were not stained in the absence of H2O2 or 3', 3'-diaminobenzidine. Histological features of mucosal inflammation, epithelium damage, and lymphoid follicle formation were graded on a scale of 0-3 for each specimen under a light microscope (magnification, × 25), and the median score was used. According to the Sydney system (27), neutrophil infiltration into the mucosa was evaluated as follows: 0, none; 1, mild; 2, moderate; and 3, severe. As described by Atherton et al. (1), epithelium damage was graded as follows: 0, no exfoliation; 1, exfoliation of <30% of epithelium; 2, 30-70% exfoliation; and 3, >70% exfoliation. Similarly, lymphoid follicle formation was also graded as follows: 0, no follicle; 1, follicles of <30% under the muscularis mucosa; 2, 30-70% follicles; and 3, >70% follicles.
Determination of viable H. pylori. Viable H. pylori in the stomach was assayed according to our previously reported method (34). The animals were killed, and their stomachs were excised. The stomachs were homogenized in 20 ml PBS. The diluted homogenates were applied onto Brucella agar (GIBCO BRL) plates supplemented with 10% horse blood (Nippon Bio-Test, Tokyo, Japan) and (in µg/ml) 2.5 amphotericin B (Sigma, St. Louis, MO), 9 vancomycin (Sigma), 0.32 polymyxin B (Sigma), 5 trimethoprim (Sigma), and 50 2,3,5-triphenyltetrazolium chloride (Wako Pure Chemicals). The plates were incubated at 37°C under a microaerophilic atmosphere for 7 days. The number of colonies was counted, and viable H. pylori was expressed as CFU per stomach.
Determination of neutrophil chemokine, interferon-, and
interleukin-10 production.
The production of cytokine-induced neutrophil chemoattractant
[CINC/KC; interleukin (IL)-8 family chemokine in rodents], interferon (IFN)-
, and IL-10 was assayed according to the method of Noach et
al. (26) with slight modifications. Gastric specimens (the fundus near the antrum) were taken from normal and H. pylori-infected animals. After being washed with PBS, the tissues
were minced and then incubated in 1 ml of Dulbecco's modified Eagle's
medium supplemented with 2.5% fetal bovine serum and the above
antibiotics at 37°C for 20 h under 5% CO2 in air.
Thereafter, the tissues were homogenized in the culture medium
containing 0.1 mM phenylmethylsulfonyl fluoride and 1 µg/ml
leupeptin. The homogenates were centrifuged at 10,000 g for
20 min. The amounts of CINC/KC, IFN-
, and IL-10 in the resulting
supernatants were determined by enzyme immunoassay (CINC/KC EIA kit;
Immuno Biological Laboratories, Fujioka, Japan) and ELISA (IFN-
ELISA kit and IL-10 ELISA kit; Biosource International, Camarillo, CA),
respectively. CINC/KC, IFN-
, and IL-10 production was expressed as
picograms per milligram tissue per 20 h.
Drugs. NS-398 (kindly synthesized by Nippon Chemiphar, Tokyo, Japan) and indomethacin (Sigma) were finely dispersed in Tween 80 (10 mg/50 µl) and then suspended by adding saline to the desired concentrations. In our preliminary experiment, NS-398 at 20 mg/kg, but not at 10 mg/kg, had inhibited PGE2 production in normal mucosa, indicating that 10 mg/kg NS-398 was ineffective in COX-1 activity. In addition, indomethacin even at 2 mg/kg had potently reduced PGE2 levels in normal mucosa, but repeated administration of 5 mg/kg indomethacin had been lethal to H. pylori-infected gerbils. Therefore, the doses of 10 mg/kg and 2 mg/kg were selected for NS-398 and indomethacin, respectively. The drugs were subcutaneously administered once daily for 4 wk from 2 wk of H. pylori infection. The animals were given the drugs in a volume of 10 ml/kg body wt. Control animals received the vehicle alone.
Statistical analysis. Data are presented as means ± SE of 3-6 animals/group. Statistical differences were evaluated by Student's t-test or Mann-Whitney U-test. P values of <0.05 were considered significant.
![]() |
RESULTS |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
COX-2 induction in the gastric mucosa by H. pylori infection. In our model of H. pylori infection, H. pylori is colonized for at least 10 mo in the gastric mucosa of all gerbils given the bacteria (20, 34). The number of viable H. pylori in the stomach reached a plateau level from 2 wk after the inoculation. In addition, gastritis with hemorrhagic mucosal lesions and gastric ulcers were generated only in the fundus near the antrum from 4 wk and 20 wk of the infection, respectively (20, 34). Thus the region is highly sensitive to H. pylori, and the following parameters, except for H. pylori viability, were measured in the region.
At first, we examined the expression of COX proteins and PGE2 production in the gastric mucosa during H. pylori infection (Fig. 1). On Western blot analysis, COX-1 protein was detected both in normal mucosa and in the mucosa with H. pylori infection. The level of COX-1 protein remained nearly constant during the infection. In contrast, COX-2 protein was not found in normal or H. pylori-infected (2 wk) mucosa, but H. pylori infection for >4 wk induced the expression of COX-2 protein. PGE2 production in normal mucosa was 36.7 ± 5.6 pg · mg
|
Effects of NS-398 and indomethacin on H. pylori-induced PGE2 production and gastritis. As shown in Fig. 1, the increase in PGE2 production was associated with COX-2 induction in the H. pylori-infected mucosa. To investigate the role of COX-2 in H. pylori-induced gastritis, we examined the effects of NSAIDs on PGE2 production and gastric pathology caused by H. pylori.
NS-398 (a COX-2-selective inhibitor) at 10 mg/kg or indomethacin (a nonselective COX inhibitor) at 2 mg/kg was administered for 4 wk to normal and H. pylori-infected (2 wk) animals. NS-398 failed to inhibit PGE2 production in normal mucosa but significantly reduced the H. pylori-increased PGE2 production (Fig. 2). In contrast, indomethacin potently inhibited PGE2 production in both normal and H. pylori-infected mucosa. Significant differences were observed between the effects of NS-398 and indomethacin in the H. pylori-infected animals.
|
|
|
Effects of NS-398 and indomethacin on H. pylori infection and the
production of CINC/KC, IFN-, and IL-10.
Since NS-398 and indomethacin aggravated H. pylori-induced
gastritis, we determined the number of viable H. pylori in
the stomach after treatment with the drugs. The number of H. pylori colonized in the stomach was 2.19 ± 0.56 × 105 CFU in the control group. Both NS-398
(1.92 ± 0.42 × 105 CFU) and indomethacin
(1.51 ± 0.38 × 105 CFU) tended to reduce H. pylori number, but there were no significant differences between
the drug-treated and control groups.
|
![]() |
DISCUSSION |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
In the present study, H. pylori infection for >4 wk
induced COX-2 expression in the gastric mucosa of Mongolian gerbils.
Recent studies with human gastric specimens also revealed that COX-2 expression is absent in normal mucosa but is profoundly induced in
H. pylori-positive gastritis (22, 31, 33). In
contrast, COX-1 protein was constitutively expressed in both normal and H. pylori-infected mucosa of gerbils. PGE2
production was elevated with COX-2 protein expression, and the
increased production was significantly inhibited by NS-398 at 10 mg/kg,
at which dose PGE2 production in normal mucosa was not
affected. These results indicate that COX-2 functionally contributes to
the increased PGE2 production in H. pylori-infected mucosa. Similarly, COX-1 also produces
PGE2 in H. pylori-infected mucosa, because 2 mg/kg indomethacin reduced PGE2 production in normal mucosa
and the inhibitory effect of 2 mg/kg indomethacin on PGE2
production in H. pylori-infected mucosa was more potent than
that of 10 mg/kg NS-398. Romano et al. (29) reported that
adhesion of H. pylori on cultured gastric cancer MKN28 cells
results in COX-2 mRNA expression. However, it is unclear whether the
direct effect of H. pylori on gastric cells is crucial for
COX-2 expression in in vivo, because COX-2 protein was not expressed
even at 2 wk of H. pylori infection in our model. COX-2
expression may result from H. pylori-associated mucosal
inflammation, because 4 wk of H. pylori infection was required for gastritis and COX-2 expression. In fact, it is also known
that H. pylori infection increases the expression of
cytokines such as IL-1 and tumor necrosis factor-, which serve as
potent COX-2 inducers (13, 23, 37). Further investigation
is needed to clarify the relationship between H. pylori
infection and COX-2 induction in in vivo.
In H. pylori infection, both bacterial and host factors are believed to contribute to gastric mucosal damage. Regarding host factors, it is suspected that inflammatory responses may be involved in H. pylori-induced gastritis (3, 5, 10, 43). Blaser (3) speculated that severe mucosal inflammation disrupts gastric epithelial functions and therefore might be deleterious to the mucosa. In the present study, NS-398 and indomethacin significantly promoted neutrophil infiltration and lymphoid follicle formation induced by H. pylori infection, and the effect of indomethacin was more potent than that of NS-398. The effects of the NSAIDs on inflammatory cells were associated with inhibition of mucosal PG production. These results indicate that both COX-1- and COX-2-derived PGs might suppress the host inflammatory response to H. pylori infection, i.e., PGs might exert anti-inflammatory effects in H. pylori-infected gastric mucosa. Consequently, aggravation of H. pylori-induced epithelial damage and mucosal erosions by the NSAIDs might result from potentiation of mucosal inflammation.
In response to H. pylori infection, mucosal defense mechanisms might be activated. Because PGs are important defensive factors in the gastric mucosa (9, 28), COX-2 induction is reasonable for protection of the mucosa against H. pylori. PGE2 increases blood flow and secretion of mucus and bicarbonate, inhibits acid secretion, and directly protects gastric cells against toxic stimuli (9). We reported that H. pylori infection for 2 and 4 wk causes an increase in mucus synthesis in the gastric mucosa of gerbils and that both NS-398 and indomethacin suppress the increased mucus synthesis (35). The decreases in these PG defensive responses to H. pylori infection are suggested to contribute to aggravation of mucosal inflammation caused by COX inhibition.
In addition, downregulation of proinflammatory cytokine expression by PGs is also likely to be important for mucosal protection against H. pylori infection. In the patients with H. pylori-positive gastritis, the IL-8 level in the gastric mucosa is significantly higher than in ones with H. pylori-negative gastritis (6, 42). Similarly, in our gerbil model, CINC/KC production was induced by H. pylori infection. NS-398 and indomethacin significantly potentiated CINC/KC production in H. pylori-infected mucosa. CINC/KC belongs to the IL-8 chemokine family and is considered to play a crucial role in neutrophil infiltration in rodents because rodent counterparts of IL-8 have not been identified (40). The increased CINC/KC production accounts for enhancement of neutrophil infiltration by the NSAIDs in H. pylori-infected mucosa. These results suggest that PGs, derived from COX-2 as well as COX-1, downregulate CINC/KC production, resulting in suppression of neutrophil infiltration in H. pylori-infected mucosa.
Gastric mucosal T-lymphocyte response to H. pylori infection
is dependent predominantly on type 1 helper T-lymphocytes, which are
characterized as IFN- secreting cells (8, 10, 19). Mohammadi et al. (25) reported that in vivo neutralization
of IFN-
by administration of anti-IFN-
antibody causes a
significant reduction of gastric inflammation induced by
Helicobacter felis in mice. Irisawa et al.
(17) reported that H. pylori-induced mucosal
damage is augmented by the overexpression of IFN-
in the stomach of
mice. Furthermore, Sawai et al. (30) reported that there
is no inflammatory pathology in the gastric mucosa of H. pylori-infected IFN-
gene-deficient mice, whereas inflammatory cell infiltration and erosions are observed in the mucosa of H. pylori-infected wild-type mice. Consequently, it has been widely accepted that IFN-
is one of the important causal factors for H. pylori-associated diseases. We also confirmed that
H. pylori infection increases IFN-
production in gerbils
and found that the increase in IFN-
production is potentiated by
NS-398 and indomethacin. Similar to the case of CINC/KC production,
these results suggest that PGs, derived from both COX-1 and COX-2,
downregulate IFN-
production, resulting in an attenuation of mucosal
inflammation induced by H. pylori infection.
In contrast, IL-10 production was induced by H. pylori
infection, as observed in human patients (42), but
was not affected by NS-398 or indomethacin. IL-10 has been shown to
inhibit the production of cytokines, including IL-8 and IFN-
(2, 7). The failure to promote IL-10 level did not
attenuate the NSAID-increased production of CINC/KC and IFN-
in
H. pylori-infected mucosa. In addition, the fact that NSAIDs
had no effect on IL-10 production suggests that the NSAID-induced
increases in CINC/KC and IFN-
levels are due to the increased
production and do not result from the inhibition of peptidase activity
during culturing.
The number of viable H. pylori in the stomach was reduced by
NS-398 and indomethacin, although there were no significant differences between the drug-treated and control groups. This result indicates that
the deleterious effects of the NSAIDs on the gastric mucosa do not
result from the increase in the number of viable H. pylori. Caselli et al. (4) reported the similar result that, in
H. pylori-positive patients with rheumatoid arthritis, the
H. pylori-positive rate is significantly reduced by NSAID
treatment compared with the nontreated group. To our knowledge, there
have been no reports that NSAIDs possess an anti-H. pylori
activity, and we confirmed that the minimal inhibitory
concentration of indomethacin toward H. pylori is 50 µg/ml
and is quite a bit higher than that of clarithromycin (0.06 µg/ml).
An attenuation of mucus synthesis may be related to the decrease in
H. pylori number. The gastric mucus layer is known to serve
as the major habitat of H. pylori, and NSAIDs inhibit the
increased mucus synthesis in H. pylori-infected mucosa of gerbils. Alternatively, the increase in IFN- production may result in the decrease in H. pylori number. In a recent study by
Sawai et al. (30), the number of H. pylori in
the stomach in IFN-
gene-deficient mice was higher than that in
wild-type mice, suggesting that IFN-
plays a protective role in
H. pylori infection. Interestingly, we found that IFN-
,
in concert with H. pylori, potently reduces mucus secretion
by cultured gastric epithelial cells (36).
COX-2-selective inhibitors are expected as new NSAIDs without ulcerogenic effect. In fact, COX-2-selective NSAIDs such as nimesulide and celecoxib are clinically used as anti-inflammatory drugs, but the incidences of their gastric side effects are significantly lower than those of conventional NSAIDs. However, our results suggest the possibility that COX-2-selective NSAIDs may have an injurious effect on the gastric mucosa of H. pylori-positive patients. Wallace et al. (39) reported that, to achieve a desirable anti-inflammatory effect, COX-2-selective NSAIDs needed to be given at high doses in which both COX-1 and COX-2 activities are inhibited. If so, the safety of COX-2-selective NSAIDs may be lowered, especially in H. pylori-infected patients. Overall, independent of type of NSAID, NSAID users should be aware of these side effects if they are infected with H. pylori. In the case of application of COX-2-selective NSAIDs to preexisting gastric ulcers, the drugs also exert an unfavorable effect. We previously reported that gastric ulcer healing is significantly impaired by NS-398 at low doses in which COX-2 activity only is inhibited in rats (32).
We conclude that COX-2 as well as COX-1 has an anti-inflammatory effect in H. pylori-induced gastritis in Mongolian gerbils. COX-2-selective inhibitors may aggravate gastritis if used in H. pylori-positive patients.
![]() |
ACKNOWLEDGEMENTS |
---|
We thank Y. Keto, A. Minamide, N. Kobayashi, Y. Naka, and S. Kitazawa for technical assistance. In addition, we are very grateful to S. Takagi (Nihon SLC, Hamamatsu, Japan) for kindly providing the Mongolian gerbils.
![]() |
FOOTNOTES |
---|
This research was supported by grants from the Ministry of Education, Science, Sports and Culture of Japan [Grant-in-Aid for Scientific Research (B) #09470508 and #11470490].
Address for reprint requests and other correspondence: S. Takahashi, Dept. of Biopharmaceutics, Kyoto Pharmaceutical Univ., Misasagi, Yamashina, Kyoto 607-8414, Japan (E-mail: takahasi{at}mb.kyoto-phu.ac.jp).
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
Received 13 March 2000; accepted in final form 11 May 2000.
![]() |
REFERENCES |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
1.
Atherton, JC,
Peek RM, Jr,
Tham KT,
Cover TL,
and
Blaser MJ.
Clinical and pathological importance of heterogeneity in vacA, the vacuolating cytotoxin gene of Helicobacter pylori.
Gastroenterology
112:
92-99,
1997[ISI][Medline].
2.
Benjamin, D,
Knobloch TJ,
and
Dayton MA.
Human B-cell interleukin-10: B-cell lines derived from patients with acquired immunodeficiency syndrome and Burkitt's lymphoma constitutively secrete large quantities of interleukin-10.
Blood
80:
1289-1298,
1992[Abstract].
3.
Blaser, MJ.
Hypotheses on the pathogenesis and natural history of Helicobacter pylori-induced inflammation.
Gastroenterology
102:
720-727,
1992[ISI][Medline].
4.
Caselli, M,
Pazzi P,
LaCorte R,
Aleotti A,
Trevisani I,
and
Stabellini G.
Campylobacter-like organisms, nonsteroidal anti-inflammatory drugs and gastric lesions in patients with rheumatoid arthritis.
Digestion
44:
101-104,
1989[ISI][Medline].
5.
Crabtree, JE.
Role of cytokines in pathogenesis of Helicobacter pylori-induced mucosal damage.
Dig Dis Sci
43:
46S-55S,
1998[ISI][Medline].
6.
Crabtree, JE,
Wyatt JI,
Trejdosiewicz LK,
Peichl P,
Nichols PH,
Ramsay N,
Primrose JN,
and
Lindley IJD
Interleukin-8 expression in Helicobacter pylori infected, normal, and neoplastic gastroduodenal mucosa.
J Clin Pathol
47:
61-66,
1994[Abstract].
7.
D'Andrea, A,
Aste-Amexaga M,
Valiante NM,
Ma X,
Kubin M,
and
Trinchieri G.
Interleukin 10 (IL-10) inhibits human lymphocyte interferon -production by suppressing natural killer cell stimulatory factor/IL-12 synthesis in accessory cells.
J Exp Med
178:
1041-1048,
1993[Abstract].
8.
D'Elios, MM,
Manghetti M,
De Carli M,
Costa F,
Baldari CT,
Burroni D,
Telford JL,
Romagnani S,
and
Del Prete G.
T helper 1 effector cells specific for Helicobacter pylori in gastric antrum of patients with peptic ulcer disease.
J Immunol
158:
962-967,
1997[Abstract].
9.
Eberhart, CE,
and
DuBois RN.
Eicosanoids and the gastrointestinal tract.
Gastroenterology
109:
285-301,
1995[ISI][Medline].
10.
Ernst, PB,
Crowe SE,
and
Reyes VE.
How does Helicobacter pylori cause mucosal damage? The inflammatory response.
Gastroenterology
113:
S35-S42,
1997[ISI][Medline].
11.
Fujita, H,
Takahashi S,
and
Okabe S.
Mechanism by which indomethacin delays the healing of acetic acid-induced ulcers in rats. Role of neutrophil antichemotactic and chemotactic activities.
J Physiol Pharmacol
49:
71-82,
1998[ISI][Medline].
12.
Gierse, JK,
Hauser SD,
Creely DP,
Koboldt C,
Rangwala SH,
Isakson PC,
and
Seibert K.
Expression and selective inhibition of the constitutive and inducible forms of human cyclo-oxygenase.
Biochem J
305:
479-484,
1995[ISI][Medline].
13.
Herschman, HR.
Prostaglandin synthase 2.
Biochim Biophys Acta
1299:
125-140,
1996[ISI][Medline].
14.
Hirata, T,
Ukawa H,
Yamakuni H,
Kato S,
and
Takeuchi K.
Cyclo-oxygenase isozymes in mucosal ulcerogenic and functional responses following barrier disruption in rat stomachs.
Br J Pharmacol
122:
447-454,
1997[Abstract].
15.
Hirayama, F,
Takagi S,
Kusuhara H,
Iwao E,
Yokoyama Y,
and
Ikeda Y.
Induction of gastric ulcer and intestinal metaplasia on Mongolian gerbils infected with Helicobacter pylori.
J Gastroenterol
31:
755-757,
1996[ISI][Medline].
16.
Ikeno, T,
Ota H,
Sugiyama A,
Ishida K,
Katsuyama T,
Genta RM,
and
Kawasaki S.
Helicobacter pylori-induced chronic active gastritis, intestinal metaplasia, and gastric ulcer in Mongolian gerbils.
Am J Pathol
154:
951-960,
1999
17.
Irisawa, A,
Saito A,
Sato Y,
Obara K,
Nishimaki T,
and
Kasukawa R.
Induction of mucosal damage of stomach by inoculation of plasmid DNA encoding interferon- (Abstract).
Gastroenterology
114:
A158,
1998[ISI].
18.
Kargman, S,
Charleson S,
Cartwright M,
Frank J,
Riendeau D,
Mancini J,
Evans J,
and
O'Neill G.
Characterization of prostaglandin G/H synthase 1 and 2 in rat, dog, monkey, and human gastrointestinal tracts.
Gastroenterology
111:
445-454,
1996[ISI][Medline].
19.
Karttunen, R,
Karttunen T,
Ekre HPT,
and
MacDonald TT.
Interferon gamma and interleukin-4 secreting cells in the gastric antrum in Helicobacter pylori positive and negative gastritis.
Gut
36:
341-345,
1995[Abstract].
20.
Keto, Y,
Takahashi S,
and
Okabe S.
Healing of Helicobacter pylori-induced gastric ulcers in Mongolian gerbils. Combined treatment with omeprazole and clarithromycin.
Dig Dis Sci
44:
257-265,
1999[ISI][Medline].
21.
Matsumoto, S,
Washizuka Y,
Matsumoto Y,
Tawara S,
Ikeda F,
Yokota Y,
and
Karita M.
Induction of ulceration and severe gastritis in Mongolian gerbils by Helicobacter pylori infection.
J Med Microbiol
46:
391-397,
1997[Abstract].
22.
McCarthy, CJ,
Crofford LJ,
Greenson J,
and
Scheiman JM.
Cyclooxygenase-2 expression in gastric antral mucosa before and after eradication of Helicobacter pylori infection.
Am J Gastroenterol
94:
1218-1223,
1999[ISI][Medline].
23.
Mitchell, JA,
Larkin S,
and
Williams TJ.
Cyclooxygenase-2: regulation and relevance in inflammation.
Biochem Pharmacol
50:
1535-1542,
1995[ISI][Medline].
24.
Mizuno, H,
Sakamoto C,
Matsuda K,
Wada K,
Uchida T,
Noguchi H,
Akamatsu T,
and
Kasuga M.
Induction of cyclooxygenase 2 in gastric mucosal lesions and its inhibition by the specific antagonist delays healing in mice.
Gastroenterology
112:
387-397,
1997[ISI][Medline].
25.
Mohammadi, M,
Czinn S,
Redline R,
and
Nedrud J.
Helicobacter-specific cell mediated immune responses display a predominant Th1 phenotype and promote a delayed-type hypersensitivity response in the stomachs of mice.
J Immunol
156:
4729-4738,
1996
26.
Noach, LA,
Bosma NB,
Jansen J,
Hoek FJ,
and
van Deventer SJH
Mucosal tumor necrosis factor-, interleukin-1
, and interleukin-8 production in patients with Helicobacter pylori infection.
Scand J Gastroenterol
29:
425-429,
1994[ISI][Medline].
27.
Price, AB.
The Sydney system: histological division.
J Gastroenterol Hepatol
6:
209-222,
1991[ISI][Medline].
28.
Robert, A,
Nezamis JE,
Lancaster C,
and
Hancher AJ.
Protection by prostaglandins in rat. Prevention of gastric necrosis produced by alcohol, HCl, hypertonic NaCl, and thermal injury.
Gastroenterology
77:
433-443,
1979[ISI][Medline].
29.
Romano, M,
Ricci V,
Memoli A,
Tuccillo C,
Di Popolo A,
Sommi P,
Acquaviva AM,
Del Vecchio Blanco C,
Bruni CB,
and
Zarrilli R.
Helicobacter pylori up-regulates cyclooxygenase-2 mRNA expression and prostaglandin E2 synthesis in MKN28 gastric mucosal cells in vitro.
J Biol Chem
273:
28560-28563,
1998
30.
Sawai, N,
Kita M,
Kodama T,
Tanahashi T,
Yamaoka Y,
Tagawa Y,
Iwakura Y,
and
Imanishi J.
Role of gamma interferon in Helicobacter pylori-induced gastric inflammatory responses in a mouse model.
Infect Immun
67:
279-285,
1999
31.
Sawaoka, H,
Kawano S,
Tsuji S,
Tsujii M,
Sun W,
Gunawan ES,
and
Hori M.
Helicobacter pylori infection induces cyclooxygenase-2 expression in human gastric mucosa.
Prostaglandins Leukot Essent Fatty Acids
59:
313-316,
1998[ISI][Medline].
32.
Shigeta, J,
Takahashi S,
and
Okabe S.
Role of cyclooxygenase-2 in the healing of gastric ulcers in rats.
J Pharmacol Exp Ther
286:
1383-1390,
1998
33.
Sidong, FU,
Ramanujam KS,
Wong A,
Fantry GT,
Drachenberg CB,
James SP,
Meltzer SJ,
and
Wilson KT.
Increased expression and cellular localization of inducible nitric oxide synthase and cyclooxygenase 2 in Helicobacter pylori gastritis.
Gastroenterology
116:
1319-1329,
1999[ISI][Medline].
34.
Takahashi, S,
Keto Y,
Fujita H,
Muramatsu H,
Nishino T,
and
Okabe S.
Pathological changes in the formation of Helicobacter pylori-induced gastric lesions in Mongolian gerbils.
Dig Dis Sci
43:
754-765,
1998[ISI][Medline].
35.
Takahashi, S,
Keto Y,
and
Okabe S.
Changes in mucus synthesis in the gastric mucosa of Mongolian gerbils during H. pylori infection (Abstract).
J Gastroenterol Hepatol
12, Suppl:
D211,
1997.
36.
Takahashi, S,
Nakamura E,
and
Okabe S.
Effects of cytokines, without and with Helicobacter pylori components, on mucus secretion by cultured gastric epithelial cells.
Dig Dis Sci
43:
2301-2308,
1998[ISI][Medline].
37.
Takahashi, S,
Shigeta J,
Inoue H,
Tanabe T,
and
Okabe S.
Localization of cyclooxygenase-2 and regulation of its mRNA expression in gastric ulcers in rats.
Am J Physiol Gastrointest Liver Physiol
275:
G1137-G1145,
1998
38.
Vane, JR.
Inhibition of prostaglandin biosynthesis as a mechanism of action of aspirin-like drugs.
Nature
231:
232-235,
1971.
39.
Wallace, JL,
Bak A,
McKnight W,
Asfaha S,
Sharkey KA,
and
MacNaughton WK.
Cyclooxygenase 1 contributes to inflammatory responses in rats and mice: implications for gastrointestinal toxicity.
Gastroenterology
115:
101-109,
1998[ISI][Medline].
40.
Watanabe, K,
Iida M,
Takaishi K,
Suzuki T,
Hamada Y,
Iizuka Y,
and
Tsurufuji S.
Chemoattractants for neutrophils in lipopolysaccharide-induced inflammatory exudate from rats are not interleukin-8 counterparts but gro-gene-product/melanoma-growth stimulating-activity-related factors.
Eur J Biochem
214:
267-270,
1993[Abstract].
41.
Watanabe, T,
Tada M,
Nagai H,
Sasaki S,
and
Nakao M.
Helicobacter pylori infection induces gastric cancer in Mongolian gerbils.
Gastroenterology
115:
642-648,
1998[ISI][Medline].
42.
Yamaoka, Y,
Kita M,
Kodama T,
Sawai N,
Kashima K,
and
Imanishi J.
Expression of cytokine mRNA in gastric mucosa with Helicobacter pylori infection.
Scand J Gastroenterol
30:
1153-1159,
1995[ISI][Medline].
43.
Yokota, K,
Kobayashi K,
Kawahara Y,
Hayashi S,
Hirai Y,
Mizuno M,
Okada H,
Akagi T,
Tsuji T,
and
Oguma K.
Gastric ulcers in SCID mice induced by Helicobacter pylori infection after transplanting lymphocytes from patients with gastric lymphoma.
Gastroenterology
117:
893-899,
1999[ISI][Medline].