title: Immunoglobulin M, C-reactive protein and complement activation in rat hepatic ischemia-reperfusion injury.
abstract: BACKGROUND
  Ischemia-reperfusion (I/R) models have shown that C-reactive protein (CRP) and immunoglobulin M (IgM) are involved in complement activation. Binding of CRP and IgM to damaged cell membranes initiates complement activation and aggravates I/R injury in various organs. However, the time course of CRP- and IgM-mediated complement activation and the relation to hepatocellular injury and inflammation in liver I/R are unknown.
  
  
  OBJECTIVE
  To evaluate the time course of IgM- and CRP-related complement activation and the relation to hepatocellular injury and inflammation in a hepatic I/R rat model.
  
  
  METHODS
  Male Wistar rats were allocated to (1) five groups of animals exposed to 60 min of partial ischemia (70%) induced via clamping of the left segmental portal triad, followed by 0, 3, 6, 12 or 24 h of reperfusion (n = 6 in each group); (2) five groups of sham-operated animals with corresponding reperfusion times (n = 5), and (3) a control group sacrificed before ischemia (n = 5). Hepatocellular injury, inflammatory response, rat plasma CRP and IgM levels and immunohistochemical depositions of CRP, IgM and C3 were assessed for each group.
  
  
  RESULTS
  Histopathological injury scores of hematoxylin and eosin sections of ischemic liver lobes demonstrated increasing values throughout the reperfusion time with a peak at 12 h. Plasma aminotransferases (alanine aminotransferase and aspartate aminotransferase) significantly increased after 3 h of reperfusion, peaking at 6 h (3,100 ± 800 U/l; p < 0.05). Hepatic neutrophil influx significantly increased from 3 to 6 h of reperfusion (p < 0.05) and demonstrated the highest value at 12 h (1.1 ± 0.2 U/mg of protein). Plasma IL-6 levels in the ischemia groups showed peak values after 6 h of reperfusion, decreasing significantly thereafter (p < 0.05). Plasma CRP values reached highest levels after 3 h of reperfusion (mean 91 ± 5% of control pool), decreasing significantly thereafter. Rat IgM concentrations in plasma did not significantly change throughout the reperfusion time. Immunohistochemical depositions of IgM, CRP and C3 in ischemic lobes demonstrated a similar pattern in time, reaching maximum values at 12 h of reperfusion. The percentages of depositions of CRP and IgM were significantly correlated [r(S) = 0.569; p < 0.001; Spearman test]. The time course of C3 and CRP depositions throughout reperfusion and C3 and IgM staining were significantly similar [r(S) = 0.797 and r(S) = 0.656, respectively; p < 0.0001; ANOVA].
  
  
  CONCLUSIONS
  CRP and IgM depositions demonstrate a parallel time course throughout the reperfusion to hepatocellular damage, inflammatory response and activated complement deposition in this rat hepatic I/R model. Furthermore, the time course of CRP and IgM depositions was significantly similar to that of activated complement depositions.
authors.affiliation: Surgical Laboratory, Department of Surgery, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: G M P
authors.initials: GM
authors.lastname: Diepenhorst
authors.forename: W
authors.initials: W
authors.lastname: de Graaf
authors.forename: H W
authors.initials: HW
authors.lastname: Niessen
authors.forename: A K
authors.initials: AK
authors.lastname: van Vliet
authors.forename: C E
authors.initials: CE
authors.lastname: Hack
authors.forename: T M
authors.initials: TM
authors.lastname: van Gulik
mesh_terms: D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D001219:Aspartate Aminotransferases / Q000097:blood; D002097:C-Reactive Protein / Q000378:metabolism*; D003167:Complement Activation; D022781:Hepatocytes / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D007075:Immunoglobulin M / Q000097:blood*; D015850:Interleukin-6 / Q000097:blood; D008099:Liver / Q000276:immunology* / Q000293:injuries* / Q000473:pathology; D008297:Male; D009504:Neutrophils / Q000276:immunology / Q000473:pathology; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000097:blood* / Q000276:immunology*
chemical_list: D007075:Immunoglobulin M; D015850:Interleukin-6; D002097:C-Reactive Protein; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase