title: Induction of inducible nitric oxide synthase in hepatocytes isolated from rats with ischemia-reperfusion injury.
abstract: BACKGROUND
  Recent evidence indicates that nitric oxide (NO) has a crucial role in hepatic ischemia-reperfusion (I/R) injury. However, little is known about how I/R influences the gene expression of inducible nitric oxide synthase (iNOS) in hepatocytes. Under inflammatory conditions, we compared the induction of iNOS in hepatocytes isolated from normal and I/R-treated rats.
  
  
  METHODS
  Hepatocytes were isolated using the collagenase perfusion method from rats treated with I/R (30-minute ischemia of middle and left lobes, followed by 3-hour reperfusion) or sham operation (control): Primary cultures of rat hepatocytes were incubated with an inflammatory cytokine, interleukin-1beta (IL-1beta), to compare the iNOS induction/NO production between the 2 groups.
  
  
  RESULTS
  Both control and I/R groups had no production of nitrite (a stable metabolite of NO) in the absence of IL-1beta. In the control group, IL-1beta stimulated dose- and time-dependent production of NO. The I/R group showed more than 2-fold increased levels of NO production. Western and Northern blot analyses revealed that the I/R group also showed increased levels of iNOS protein and its messenger RNA.
  
  
  CONCLUSIONS
  These results suggest that I/R directly affects the inducibility of the iNOS gene in hepatocytes by IL-1beta. Increased NO may be associated with protective or toxic effects in hepatic I/R injury.
authors.affiliation: Department of Surgery, Kansai Medical University, Osaka, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Yanagida
authors.forename: M
authors.initials: M
authors.lastname: Kaibori
authors.forename: M
authors.initials: M
authors.lastname: Yamada
authors.forename: K
authors.initials: K
authors.lastname: Habara
authors.forename: N
authors.initials: N
authors.lastname: Yokoigawa
authors.forename: A-H
authors.initials: AH
authors.lastname: Kwon
authors.forename: Y
authors.initials: Y
authors.lastname: Kamiyama
authors.forename: T
authors.initials: T
authors.lastname: Okumura
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004790:Enzyme Induction; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D022781:Hepatocytes / Q000201:enzymology*; D007375:Interleukin-1 / Q000494:pharmacology; D008102:Liver Circulation; D008297:Male; D019001:Nitric Oxide Synthase / Q000096:biosynthesis / Q000235:genetics*; D052247:Nitric Oxide Synthase Type II; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000201:enzymology* / Q000473:pathology
chemical_list: D007375:Interleukin-1; D012333:RNA, Messenger; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat