title: Epimorphin, a morphogenic protein, induces proteases in rodent hepatocytes through NF-kappaB.
abstract: OBJECTIVE
  Epimorphin, expressed by hepatic stellate cells in the liver, directs normal morphogenesis in various organs. The aim of this study was to clarify the mechanism by which epimorphin functions as a morphogen in vitro.
  
  
  METHODS
  Male Balb/c mice and Sprague-Dawley rats were used. First, we explored the relationship between epimorphin expression and distribution of protease-positive cells in carbon tetrachloride-induced acute liver injury. We then examined protease levels in cultured hepatocytes and signal transduction of epimorphin. Finally, we determined the requirement for proteases and NF-kappaB in spheroid formation induced by epimorphin.
  
  
  RESULTS
  Epimorphin expression was enhanced in injured areas during late recovery phase, in which protease-positive hepatocytes were localized adjacent to epimorphin-expressing cells. In vitro, epimorphin induced matrix metalloproteinase (MMP) 9, MMP 3 and urokinase type plasminogen activator (uPA) in hepatocytes. NF-kappaB mediated these protease expressions in hepatocytes. These proteases were required for epimorphin-induced and Matrigel induced spheroid. An epimorphin-neutralizing antibody also blocked spheroid formation on Matrigel, which contained epimorphin. In addition, NF-kappaB activation was also required for spheroid formation.
  
  
  CONCLUSIONS
  Epimorphin elicits hepatocyte spheroids by inducing proteases in rodent hepatocytes through NF-kappaB.
authors.affiliation: Department of Gastroenterology, Akita University School of Medicine, 1-1-1 Hondo, Akita 010-8543, Japan.
authors.forename: Kouichi
authors.initials: K
authors.lastname: Miura
authors.forename: Ryutaro
authors.initials: R
authors.lastname: Yoshino
authors.forename: Yohei
authors.initials: Y
authors.lastname: Hirai
authors.forename: Takashi
authors.initials: T
authors.lastname: Goto
authors.forename: Shigetoshi
authors.initials: S
authors.lastname: Ohshima
authors.forename: Ken-Ichiro
authors.initials: K
authors.lastname: Mikami
authors.forename: Kazuo
authors.initials: K
authors.lastname: Yoneyama
authors.forename: Daisuke
authors.initials: D
authors.lastname: Watanabe
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Sato
authors.forename: Haruki
authors.initials: H
authors.lastname: Senoo
authors.forename: Yuzo
authors.initials: Y
authors.lastname: Kodama
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Osawa
authors.forename: David A
authors.initials: DA
authors.lastname: Brenner
authors.forename: Sumio
authors.initials: S
authors.lastname: Watanabe
mesh_terms: D000818:Animals; D002251:Carbon Tetrachloride; D018920:Coculture Techniques; D005786:Gene Expression Regulation; D022781:Hepatocytes / Q000201:enzymology*; D008099:Liver / Q000166:cytology; D008103:Liver Cirrhosis / Q000139:chemically induced / Q000201:enzymology / Q000473:pathology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D016328:NF-kappa B / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction; D018874:Spheroids, Cellular; D050827:Syntaxin 1 / Q000502:physiology*
chemical_list: D016328:NF-kappa B; D050827:Syntaxin 1; D002251:Carbon Tetrachloride; D010447:Peptide Hydrolases