title: Roxithromycin inhibits nuclear factor kappaB signaling and endoplasmic reticulum stress in intestinal epithelial cells and ameliorates experimental colitis in mice.
abstract: Roxithromycin is known to have anti-inflammatory and immunoregulatory activity. However, little information is available on the effect of roxithromycin in intestinal inflammation. The aim of this study was to investigate the effect of roxithromycin on NF- κB signaling and ER stress in intestinal epithelial cells (IECs) and the effect of roxithromycin on dextran sulfate sodium (DSS)-induced acute colitis in a murine model. HCT116 cells and COLO205 cells were pretreated with roxithromycin and then stimulated with tumor necrosis factor-α (TNF-α). Interleukin (IL)-8 expression was determined by real-time reverse transcription-polymerase chain reaction. Nuclear factor kappaB (NF-κB) DNA-binding activity and IκB phosphorylation/degradation were evaluated by electrophoretic mobility shift assay and Western blot analysis. The molecular markers of endoplasmic reticulum stress, including p-JNK, phosphorylated eukaryotic initiation factor 2 (p-eIF2α), C/EBP homologous protein (CHOP), and X-box binding protein 1 (XBP1) were evaluated using western blotting and PCR. Mice were given 4% DSS for five days with or without roxithromycin. Primary IECs were isolated from mice with DSS-induced colitis. Roxithromycin significantly inhibited the upregulated expression of IL-8. Pretreatment with roxithromycin markedly attenuated NF-κB DNA-binding activity and IκB phosphorylation/degradation. CHOP and XBP1 mRNA expression were enhanced in the presence of TNF-α, and it was dampened by pretreatment of roxithromycin. c-Jun-N-terminal kinase (JNK) phosphorylation and the level of p-eIF2α were also downregulated by the pretreatment of roxithromycin. Roxithromycin significantly reduced the severity of DSS-induced murine colitis, as assessed by the disease activity index, colon length, and histology. In addition, the DSS-induced phospho-IκB kinase activation was significantly decreased in roxithromycin-pretreated mice. Finally, IκB degradation was reduced in primary IECs from mice treated with roxithromycin. These results suggest that roxithromycin may have potential usefulness in the treatment of inflammatory bowel disease.
authors.affiliation: Department of Internal Medicine, Seoul National University Boramae Hospital, Seoul National University College of Medicine, Seoul 156-707, Korea.
authors.forename: Younjeong
authors.initials: Y
authors.lastname: Choi
authors.affiliation: Department of Internal Medicine, Seoul National University Boramae Hospital, Seoul National University College of Medicine, Seoul 156-707, Korea.
authors.forename: Seong-Joon
authors.initials: SJ
authors.lastname: Koh
authors.affiliation: Department of Internal Medicine, Seoul National University Boramae Hospital, Seoul National University College of Medicine, Seoul 156-707, Korea.
authors.forename: Hee Sook
authors.initials: HS
authors.lastname: Lee
authors.affiliation: Department of Internal Medicine, Seoul National University Boramae Hospital, Seoul National University College of Medicine, Seoul 156-707, Korea giwkim@hanmail.net.
authors.forename: Ji Won
authors.initials: JW
authors.lastname: Kim
authors.affiliation: Department of Internal Medicine, Seoul National University Boramae Hospital, Seoul National University College of Medicine, Seoul 156-707, Korea.
authors.forename: Byeong
authors.initials: B
authors.lastname: Gwan Kim
authors.affiliation: Department of Internal Medicine, Seoul National University Boramae Hospital, Seoul National University College of Medicine, Seoul 156-707, Korea.
authors.forename: Kook Lae
authors.initials: KL
authors.lastname: Lee
authors.affiliation: Department of Internal Medicine, Liver Research Institute, Seoul National University College of Medicine, Seoul 110-744, Korea.
authors.forename: Joo Sung
authors.initials: JS
authors.lastname: Kim
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology* / Q000627:therapeutic use; D003092:Colitis / Q000188:drug therapy* / Q000503:physiopathology; D004195:Disease Models, Animal; D024202:Electrophoretic Mobility Shift Assay; D004721:Endoplasmic Reticulum / Q000187:drug effects* / Q000502:physiology; D045325:HCT116 Cells / Q000187:drug effects / Q000502:physiology; D006801:Humans; D016209:Interleukin-8 / Q000502:physiology; D007413:Intestinal Mucosa / Q000187:drug effects* / Q000503:physiopathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D016328:NF-kappa B / Q000037:antagonists & inhibitors*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015575:Roxithromycin / Q000494:pharmacology* / Q000627:therapeutic use; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology
chemical_list: D000893:Anti-Inflammatory Agents; D016209:Interleukin-8; D016328:NF-kappa B; D015575:Roxithromycin
keywords: Roxithromycin; colitis; endoplasmic reticulum stress; nuclear factor-KappaB