title: Lutein protects against methotrexate-induced and reactive oxygen species-mediated apoptotic cell injury of IEC-6 cells.
abstract: OBJECTIVE
  High-dose chemotherapy using methotrexate (MTX) frequently induces side effects such as mucositis that leads to intestinal damage and diarrhea. Several natural compounds have been demonstrated of their effectiveness in protecting intestinal epithelial cells from these adverse effects. In this paper, we investigated the protection mechanism of lutein against MTX-induced damage in IEC-6 cells originating from the rat jejunum crypt.
  
  
  METHODS
  The cell viability, induced-apoptosis, reactive oxygen species (ROS) generation, and mitochondrial membrane potential in IEC-6 cells under MTX treatment were examined in the presence or absence of lutein. Expression level of Bcl2, Bad and ROS scavenging enzymes (including SOD, catalase and Prdx1) were detected by quantitative RT-PCR.
  
  
  RESULTS
  The cell viability of IEC-6 cells exposed to MTX was decreased in a dose- and time-dependent manner. MTX induces mitochondrial membrane potential loss, ROS generation and caspase 3 activation in IEC-6 cells. The cytotoxicity of MTX was reduced in IEC-6 cells by the 24 h pre-treatment of lutein. We found that pre-treatment of lutein significantly reduces MTX-induced ROS and apoptosis. The expression of SOD was up-regulated by the pre-treatment of lutein in the MTX-treated IEC-6 cells. These results indicated that lutein can protect IEC-6 cells from the chemo-drugs induced damage through increasing ROS scavenging ability.
  
  
  CONCLUSIONS
  The MTX-induced apoptosis of IEC-6 cells was shown to be repressed by the pre-treatment of lutein, which may represent a promising adjunct to conventional chemotherapy for preventing intestinal damages.
authors.affiliation: Division of Pediatric Surgery, Department of Surgery, Shin-Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan ; School of Medicine, Fu-Jen Catholic University, New Taipei, Taiwan.
authors.forename: Chi-Jen
authors.initials: CJ
authors.lastname: Chang
authors.forename: Ji-Fan
authors.initials: JF
authors.lastname: Lin
authors.forename: Hsun-Hsien
authors.initials: HH
authors.lastname: Chang
authors.forename: Gon-Ann
authors.initials: GA
authors.lastname: Lee
authors.forename: Chi-Feng
authors.initials: CF
authors.lastname: Hung
mesh_terms: D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D002374:Catalase / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D019610:Cytoprotection; D004789:Enzyme Activation; D004847:Epithelial Cells / Q000187:drug effects / Q000502:physiology; D007413:Intestinal Mucosa / Q000166:cytology; D014975:Lutein / Q000494:pharmacology*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008727:Methotrexate / Q000633:toxicity*; D018384:Oxidative Stress; D054464:Peroxiredoxins / Q000378:metabolism; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000378:metabolism
chemical_list: D000964:Antimetabolites, Antineoplastic; D017382:Reactive Oxygen Species; C489898:Prdx1 protein, rat; D054464:Peroxiredoxins; D002374:Catalase; D013482:Superoxide Dismutase; C505407:CASP3 protein, human; D053148:Caspase 3; D014975:Lutein; D008727:Methotrexate
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