Effects of rebamipide on bile acid-induced inhibition of gastric epithelial repair in a rabbit cell culture model.
BACKGROUND
Anti-ulcer agents exert various functional effects on gastric epithelial cells.


OBJECTIVE
The effects of a novel gastro-cytoprotective agent (rebamipide) on epithelial restoration following bile acid damage were assessed using primary cultured rabbit gastric epithelial cells.


METHODS
Rebamipide was added to complete confluent cell sheets with deoxycholic acid just after creating a cell-free wound (2 mm2). The restoration was monitored and analysed by phase contrast microscopy and an image analyser for 48 h. The migration speed was measured during the initial 3 h after wounding. Cell proliferation was detected by staining for bromodeoxyuridine (BrdU) at 12-h intervals. The labelling index was calculated per unit area. The major cytoskeletal protein actin was detected by immunohistochemical staining.


RESULTS
In the controls, restoration was completed 48 h following wounding. Deoxycholic acid retarded this process. The addition of rebamipide to deoxycholic acid abolished the bile acid-induced retardation. The migration speed was 26 microns/h in the controls. 15 microns/h in the deoxycholic acid group and 27 microns/h in the deoxycholic acid plus rebamipide group. In the controls, BrdU-positive cells, which were rarely detected in the initial 24 h, were maximal at 36 h (labelling index 1.7%). In the deoxycholic acid group, proliferation was inhibited (peak labeling index; 0.5% at 48 h). Actin-containing stress fibres were detected throughout the cells and the periphery of the lamellipodia in the controls, and were disrupted in the deoxycholic acid-treated group. Rebamipide prevented these effects.


CONCLUSIONS
Deoxycholic acid significantly retarded restoration by the inhibition of both cell migration and proliferation, potentially through an effect on the cytoskeleton. Rebamipide protected the mucosal cells from bile acid mediated injury.
Watanabe
S
S
Department of Gastroenterology, Juntendo University School of Medicine, Tokyo, Japan.
Wang
X E
XE
Hirose
M
M
Yoshizawa
T
T
Iwazaki
R
R
Oide
H
H
Kitamura
T
T
Miwa
H
H
Miyazaki
A
A
Sato
N
N
D000199:Actins / Q000032:analysis; D000409:Alanine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D000897:Anti-Ulcer Agents / Q000494:pharmacology*; D001973:Bromodeoxyuridine / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D003840:Deoxycholic Acid / Q000633:toxicity; D004195:Disease Models, Animal; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D005753:Gastric Mucosa / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D015363:Quinolones / Q000494:pharmacology*; D011817:Rabbits; D013194:Staining and Labeling / Q000379:methods; D013272:Stomach Diseases / Q000139:chemically induced / Q000188:drug therapy* / Q000473:pathology
D000199:Actins; D000897:Anti-Ulcer Agents; D015363:Quinolones; D003840:Deoxycholic Acid; D001973:Bromodeoxyuridine; C052785:rebamipide; D000409:Alanine