title: Heat-shock protein 70-overexpressing gastric epithelial cells are resistant to indomethacin-induced apoptosis.
abstract: OBJECTIVE
  Protecting intestinal mucosa from nonsteroidal anti-inflammatory drugs is still an unsolved problem. It has been revealed that apoptosis in epithelial cells as a result of mitochondrial injury is an important pathogenesis in indomethacin-induced gastric mucosal injury. In this study, we revealed the effect of overexpressed heat-shock protein 70 (HSP70) in indomethacin-induced apoptosis and oxidative stress.
  
  
  METHODS
  HSP70-overexpressing rat gastric mucosal cells (7018-RGM-1 cells) and control cells (pBK-CMV-12 cells) were used and treated with 0-500 microM of indomethacin for 24 h. Cell viability and cytotoxity were measured by a WST-8 assay and a lactate dehydrogenase release assay, respectively. Apoptosis was observed by fluorescence microscopy staining with Hoechst 33342 and propidium iodide. The expression of Bcl-2 family proteins, activation of caspase-3, and 4-hydroxy-2-nonenal (4-HNE)-modified proteins were assessed by Western blot analysis.
  
  
  RESULTS
  Indomethacin caused apoptosis of gastric epithelial cells. The 7018-RGM-1 cells survived significantly after indomethacin treatment compared to the control cells. The increase in pro-apoptotic Bad proteins, the decrease in anti-apoptotic Bcl-2 proteins, and caspase activation were all suppressed in the 7018-RGM-1 cells. A lower level of indomethacin-induced 4-HNE-modification was detected in the 7018-RGM-1 cells than in the control cells.
  
  
  CONCLUSIONS
  Overexpressed HSP70 may potentiate resistance to apoptosis and oxidative stress in indomethacin-induced gastric epithelial cell injury.
authors.affiliation: Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kamigyo-ku, Kyoto, Japan.
authors.forename: Ikuhiro
authors.initials: I
authors.lastname: Hirata
authors.forename: Yuji
authors.initials: Y
authors.lastname: Naito
authors.forename: Osamu
authors.initials: O
authors.lastname: Handa
authors.forename: Natsuko
authors.initials: N
authors.lastname: Hayashi
authors.forename: Katsura
authors.initials: K
authors.lastname: Mizushima
authors.forename: Satoko
authors.initials: S
authors.lastname: Adachi
authors.forename: Tatsushi
authors.initials: T
authors.lastname: Omatsu
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Okayama
authors.forename: Etsuko
authors.initials: E
authors.lastname: Kishimoto
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ichikawa
authors.forename: Tomohisa
authors.initials: T
authors.lastname: Takagi
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kokura
authors.forename: Michiro
authors.initials: M
authors.lastname: Otaka
authors.forename: Toshikazu
authors.initials: T
authors.lastname: Yoshikawa
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000009:adverse effects* / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism; D005753:Gastric Mucosa / Q000187:drug effects / Q000378:metabolism* / Q000503:physiopathology; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis*; D007213:Indomethacin / Q000009:adverse effects* / Q000494:pharmacology; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D051381:Rats
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D018840:HSP70 Heat-Shock Proteins; D007213:Indomethacin