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. 2000 Dec;47(6):762–770. doi: 10.1136/gut.47.6.762

Cyclooxygenase (COX) 1 and 2 in normal, inflamed, and ulcerated human gastric mucosa

L Jackson 1, K Wu 1, Y Mahida 1, D Jenkins 1, C Hawkey 1
PMCID: PMC1728133  PMID: 11076873

Abstract

BACKGROUND AND AIMS—Constitutive cyclooxygenase (COX) 1 is believed to mediate prostaglandin dependent gastric protection. However, gastric mucosa contains cells capable of expressing inducible COX-2. We therefore investigated COX-1 and COX-2 expression, localisation, and activity in normal and abnormal human gastric mucosa.
METHODS—COX-1 and COX-2 distribution was investigated by light and electron microscopic immunohistochemistry and by western blot analysis, and their contribution to prostaglandin (PG)E2 synthesis using selective enzyme inhibitors.
RESULTS—There was strong parietal cell COX-1 and COX-2 immunoreactivity in all sections and isolated cells, with macrophage and myofibroblast reactivity in some sections. Immunostaining was specifically abolished by antigen absorption. Western blot analysis confirmed COX-1 and 2 expression. COX-1 and COX-2 immunostaining was increased in Helicobacter pylori gastritis, particularly the mid glandular zone and lamina propria inflammatory cells. This was associated with increased ex vivo PGE2 synthesis (62.4 (13.5) pg/mg v 36.3 (15.5) pg/mg in uninflamed mucosa; p=0.017) which was significantly inhibited by COX-1 but not COX-2 inhibition. Increased COX-2 immunostaining in macrophages, endothelial cells, and myofibroblasts (with reduced epithelial expression) was seen at the rim of ulcers.
CONCLUSION—COX-2, as well as COX-1, is expressed by normal human gastric mucosa and is increased at the rim of ulcers. Although both are increased with H pylori, COX-1 contributes more than COX-2 to gastric PGE2 production.


Keywords: stomach; gastric mucosa; cyclooxygenases; Helicobacter pylori; ulceration; prostaglandins

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Figure 1  .

Figure 1  

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Immunohistochemistry of cyclooxygenase (COX)-1 and COX-2 enzymes in normal human gastric mucosa. Immunoperoxidase activity was positive in glandular epithelial cells when incubated with antibodies against COX-1 (A) and COX-2 (B), but negative when incubated with an anti-COX-1 antibody preabsorbed with purified COX-1 (C) and an anti-COX-2 antibody preabsorbed with purified COX-2 (D). Preincubation of COX-1 antibody with COX-2 antigen (E) and COX-2 antibody with COX-1 antigen (F) did not block staining. Original magnification: ×25 for A, C, E; ×160 for B, D, F (note, sections are not contiguous).

Figure 2  .

Figure 2  

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Immunohistochemical analysis of human gastric mucosa with antibodies to cyclooxygenase (COX)-1 (A) and human milk fat globule 2 (HMFG2) (B). Immunopositive cells are represented with morphological features of parietal cells (arrows). In cytospin preparations, cells with the morphological features of parietal cells demonstrated cytoplasmic immunoreactivity for HMFG2 (C), COX-1 (D), and COX-2 (E).

Figure 3  .

Figure 3  

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Electron photomicrography of human gastric glands stained with antibodies to cyclooxygenase (COX)-1 (A) and COX-2 (B). Immunoreactivity was localised to intracellular membrane structures (arrows), likely to be smooth endoplasmic reticulum or canicular membranes. (C, D) Western blot analysis of COX-1 and COX-2 proteins in human gastric epithelial cells. Cell lysate (10 µg, lane 4) was separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, transferred to nitrocellulose membranes, and immunoblotted with (C) anti-COX-1 and (D) anti-COX-2 antibodies. Purified COX-1 and COX-2 standards (0.5 µg) were treated in the same way as the cell lysate and are shown on lane 2 and lane 3, respectively. Molecular weight marker for protein is shown on lane 1 with 60 kDa and 30 kDa bands present. An estimated 72 kDa protein band is seen on lanes with either COX standard or epithelial cells.

Figure 4  .

Figure 4  

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Immunohistochemistry of H pylori associated gastritis showed an increase in the relative intensity of staining of the proliferative zone of the epithelium with cyclooxygenase (COX)-1 (A) and COX-2 (B) antibodies. There was also an increase in the number of cells of the lamina propria which stained positively with COX-1 (A) and COX-2 (B) antibodies.

Figure 5  .

Figure 5  

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(A) Dose dependent inhibition of prostaglandin E2 (PGE2) production by cyclooxygenase (COX)-1 inhibitor SC58560 and COX-2 inhibitor NS-398 at concentrations of 10−7 M and 10−5 M. (B) COX-1 and COX-2 inhibition of ex vivo PGE2 production from whole biopsies by SC58560 and NS -398 at concentrations of 10−5 M in the presence or absence of H pylori. *p=0.017, **p=0.002, (*)p=0.13. (C) PGE2 production by parietal cell enriched fraction of human gastric epithelial cells (control) and its inhibition by SC58560 and NS-398 at concentrations of 10−6 M and 10−8 M.

Figure 6  .

Figure 6  

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Immunohistochemistry of cyclooxygenase (COX)-2 enzyme in ulcerated human gastric mucosa. Immunoreactivity is seen on (A) macrophages in lamina propria proximal to the ulcer rim, (B) myofibroblasts at the ulcer base or granulation tissue, and (C) vascular endothelial cells in the submucosa next to the ulcer.

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