Proteome analysis of rat hepatic stellate cells

Authors

  • Dan Bach Kristensen,

    1. Hiroshima Tissue Regeneration Project, Hiroshima Prefecture Joint-Research Project for Regional Intensive, JST, Hiroshima
    2. Department of Biological Science, Graduate School of Science, Hiroshima University, Hiroshima
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  • Norifumi Kawada,

    1. Third Department of Internal Medicine, Osaka City University Medical School, Osaka, Japan
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  • Kunihiko Imamura,

    1. Hiroshima Tissue Regeneration Project, Hiroshima Prefecture Joint-Research Project for Regional Intensive, JST, Hiroshima
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  • Yuka Miyamoto,

    1. Hiroshima Tissue Regeneration Project, Hiroshima Prefecture Joint-Research Project for Regional Intensive, JST, Hiroshima
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  • Chise Tateno,

    1. Hiroshima Tissue Regeneration Project, Hiroshima Prefecture Joint-Research Project for Regional Intensive, JST, Hiroshima
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  • Shuichi Seki,

    1. Third Department of Internal Medicine, Osaka City University Medical School, Osaka, Japan
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  • Tetsuo Kuroki,

    1. Third Department of Internal Medicine, Osaka City University Medical School, Osaka, Japan
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  • Katsutoshi Yoshizato Ph.D.

    Corresponding author
    1. Hiroshima Tissue Regeneration Project, Hiroshima Prefecture Joint-Research Project for Regional Intensive, JST, Hiroshima
    2. Department of Biological Science, Graduate School of Science, Hiroshima University, Hiroshima
    • Developmental Biology Laboratory, Department of Biological Science, Graduate School of Science, Hiroshima University, 1-3-1, Kagamiyama, Higashihiroshima, Hiroshima 739-8526, Japan. fax: (81) 824 24 1492.
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Abstract

Proteome analysis was performed on cellular and secreted proteins of normal (quiescent) and activated rat hepatic stellate cells. The stellate cells were activated either in vitro by cultivating quiescent stellate cells for 9 days or in vivo by injecting rats with carbon tetrachloride for 8 weeks. A total of 43 proteins/polypeptides were identified, which altered their expression levels when the cells were activated in vivo and/or in vitro. Twenty-seven of them showed similar changes in vivo and in vitro, including up-regulated proteins such as calcyclin, calgizzarin, and galectin-1 as well as down-regulated proteins such as liver carboxylesterase 10 and serine protease inhibitor 3. Sixteen of them showed different expression levels between in vivo and in vitro activated stellate cells. These results were reproducibly obtained in 3 independent experiments. The up-regulation of calcyclin, calgizzarin, and galectin-1, as well as the down-regulation of liver carboxylesterase 10 were directly confirmed in fibrotic liver tissues. Northern blots confirmed up-regulation of the messenger RNAs (mRNAs) of calcyclin, calgizzarin, and galectin-1 in activated stellate cells, indicating that these changes were controlled at the mRNA level. In addition a list compiling over 150 stellate cell proteins is presented. The data presented here thus provide a significant new protein-level insight into the activation of hepatic stellate cells, a key event in liver fibrogenesis.

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