title: Rebamipide significantly inhibits indomethacin-induced mitochondrial damage, lipid peroxidation, and apoptosis in gastric epithelial RGM-1 cells.
abstract: Nonsteroidal antiinflammatory drugs (NSAIDs) cause complications such as gastrointestinal injury. NSAIDs were recently reported to cause mitochondrial injury: to dissipate the mitochondrial transmembrane potential (MTP), and to induce mitochondrial permeability transition pore (PTP), which liberates cytochrome c. This enzyme generates reactive oxygen species (ROS) thereby triggers caspase cascade and cellular lipid peroxidation, resulting in cellular apoptosis. However, the mechanism of this NSAID-induced MTP's role in cellular apoptosis remains unknown. Rebamipide, an antiulcer drug, is reported to scavenge ROS and to show the protective effects on indomethacin-induced tissue peroxidations. Since cytochrome c and its generation of ROS are involved in indomethacin-induced cellular apoptosis, rebamipide may attenuate mitochondrial damage. The aim of this study was to elucidate whether indomethacin induces both the MTP decrease and cellular apoptosis, and the effect of rebamipide on these phenomena. We examined the effect of rebamipide on 1) MTP change, 2) lipid peroxidation, 3) apoptosis, and 4) caspase activation using gastric mucosal epithelial cell-line treated with indomethacin. With a specially designed fluorescence analyzing microscope system, MTP change, cellular lipid peroxidation, and cellular apoptosis were investigated with the small star, filled following fluorescent dyes, MitoRed, DPPP, and Hoechst 33,258, respectively. Indomethacin treatment decreased MTP but increased both cellular lipid peroxidation and cellular apoptosis via caspase 3 and 9 activation. Rebamipide clearly inhibited these phenomena {in vitro}. We demonstrated that fluorescent dyes such as MitoRed, DPPP, and Hoechst 33,258 are useful indicators for detecting oxidative cellular injuries in living cells. Rebamipide exerts a protective effect on mitochondrial membrane stability in gastric epithelial cells.
authors.affiliation: Division of Gastroenterology, Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Ten-nohdai, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Nagano
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Matsui
authors.forename: Mutsumi
authors.initials: M
authors.lastname: Muramatsu
authors.forename: Osamu
authors.initials: O
authors.lastname: Shimokawa
authors.forename: Takeshi
authors.initials: T
authors.lastname: Shibahara
authors.forename: Akinori
authors.initials: A
authors.lastname: Yanaka
authors.forename: Akira
authors.initials: A
authors.lastname: Nakahara
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Matsuzaki
authors.forename: Naomi
authors.initials: N
authors.lastname: Tanaka
authors.forename: Yukio
authors.initials: Y
authors.lastname: Nakamura
mesh_terms: D000409:Alanine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000009:adverse effects*; D000897:Anti-Ulcer Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D018929:Cell Culture Techniques; D005456:Fluorescent Dyes; D005753:Gastric Mucosa / Q000166:cytology* / Q000473:pathology*; D007213:Indomethacin / Q000009:adverse effects*; D015227:Lipid Peroxidation; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008928:Mitochondria / Q000187:drug effects* / Q000473:pathology*; D015363:Quinolones / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D000897:Anti-Ulcer Agents; D005456:Fluorescent Dyes; D015363:Quinolones; D017382:Reactive Oxygen Species; C052785:rebamipide; D000409:Alanine; D007213:Indomethacin