Endogenous heme oxygenase induction is a critical mechanism attenuating apoptosis and restoring microvascular perfusion following limb ischemia/reperfusion

Surgery. 2004 Jul;136(1):67-75. doi: 10.1016/j.surg.2003.11.002.


Background: A protective role for endogenous heme oxygenase (HO) in the initiation of remote liver injury after limb ischemia/reperfusion has been established. This study expands on our previous work by investigating the role of endogenous HO on hepatocellular injury, hepatocyte death (necrotic and apoptotic), and microvascular perfusion at protracted post-reperfusion times.

Methods: Remote liver injury was studied after 1 hour of bilateral hind limb ischemia and 3, 6, or 24 hours of reperfusion in male C57BL6 mice. Inhibition of HO was achieved with the use of chromium mesoporphrin (CrMP). Established intravital videomicroscopy techniques were used to evaluate microvascular perfusion and hepatocyte death. Hepatocellular injury was quantified by serum alanine transaminase. Apoptosis was measured by using DNA laddering, Cell Death ELISA, and caspase-3 activity.

Results: Although significant perfusion deficits and hepatocellular injury/death occurred after 3 hours, progression of hepatocellular death beyond 6 hours was not observed. A transient increase in apoptosis was observed at 6 hours. By 24 hours, microvascular perfusion was completely restored. This lack of progression correlated with increased HO activity, observed throughout the protocol. Administration of CrMP reduced HO activity to sham nonstressed levels, and caused increased microvascular perfusion deficits, hepatocellular injury, and hepatocyte death over 24 hours. The transient increase in apoptosis was increased in duration and magnitude in CrMP-treated animals.

Conclusions: These results suggest that endogenous HO activity prevents the progression of remote liver injury after limb ischemia/reperfusion.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Apoptosis / physiology
  • Cell Death / drug effects
  • Cell Death / physiology
  • Enzyme Inhibitors / pharmacology
  • Heme Oxygenase (Decyclizing) / antagonists & inhibitors
  • Heme Oxygenase (Decyclizing) / drug effects
  • Heme Oxygenase (Decyclizing) / physiology*
  • Hepatocytes / drug effects
  • Hepatocytes / physiology
  • Hindlimb / blood supply
  • Liver Diseases / etiology*
  • Male
  • Mesoporphyrins / pharmacology
  • Mice
  • Microcirculation / drug effects
  • Microcirculation / physiopathology*
  • Reperfusion Injury / complications
  • Reperfusion Injury / physiopathology*


  • Enzyme Inhibitors
  • Mesoporphyrins
  • chromium mesoporphyrin
  • Heme Oxygenase (Decyclizing)