Protective effects of HO-1 pathway on lung injury subsequent to limb ischemia reperfusion

Kaohsiung J Med Sci. 2019 Jul;35(7):417-424. doi: 10.1002/kjm2.12070. Epub 2019 Apr 12.

Abstract

Limb ischemia reperfusion (LIR) can activate endogenous cytoprotective mechanisms by generating specific proteins against reperfusion injury in remote organs. The present study investigated the roles of heme oxygenase-1 (HO-1) pathway and the molecular mechanisms underlying the regulation of this pathway on lung injury following LIR. LIR was induced by ischemia for 4 hours followed by reperfusion for 6 hours (LIR 6 hours) or 16 hours (LIR 16 hours) in male Sprague-Dawley rats. HO-1 inducer cobalt protoporphyrin (Copp) or HO-1 inhibitor zinc protoporphyrin (Znpp) was intravenously injected 24 hours before ischemia. The animals were randomly divided into nine groups, including normal control, LIR 6 hours, LIR 16 hours, Copp, Copp + LIR 6 hours, Copp + LIR 16 hours, and Znpp, Znpp+ LIR 6 hours, and Znpp + LIR 16 hours groups (each group included four samples). Lung injury was examined through histopathology. Quantitative real-time PCR, immunohistochemistry and Western blot were applied to detect the mRNA and protein levels of HO-1, Nrf2, and Bach1. Our study showed that LIR induced Nrf2 upregulation but Bach1 downregulation to promote HO-1 expression in lung tissues. Activation of HO-1 pathway by Copp potentially enhanced Nrf2 expression but inhibition of the pathway by Znpp promoted Bach1 expression. Inducer of HO-1 pathway, Copp injection improved the lung injury. Nevertheless, Znpp injection aggravated the lung injury following LIR. Our findings suggested that activated HO-1 pathway might exert protective effects on the lung injury following LIR.

Keywords: BTB and CNC homology 1; heme oxygenase-1; limb ischemia reperfusion; lung injury; nuclear factor erythroid 2-related factor.

MeSH terms

  • Animals
  • Basic-Leucine Zipper Transcription Factors / genetics
  • Basic-Leucine Zipper Transcription Factors / metabolism
  • Enzyme Activators / pharmacology
  • Enzyme Inhibitors / pharmacology
  • Gene Expression Regulation
  • Heme Oxygenase (Decyclizing) / antagonists & inhibitors
  • Heme Oxygenase (Decyclizing) / genetics*
  • Heme Oxygenase (Decyclizing) / metabolism
  • Hindlimb / blood supply
  • Hindlimb / drug effects
  • Hindlimb / enzymology
  • Lung
  • Lung Injury / etiology
  • Lung Injury / genetics*
  • Lung Injury / pathology
  • Lung Injury / prevention & control
  • Male
  • NF-E2-Related Factor 2 / genetics
  • NF-E2-Related Factor 2 / metabolism
  • Protoporphyrins / pharmacology
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Reperfusion Injury / complications
  • Reperfusion Injury / genetics*
  • Reperfusion Injury / pathology
  • Reperfusion Injury / prevention & control
  • Signal Transduction / genetics*

Substances

  • Basic-Leucine Zipper Transcription Factors
  • Enzyme Activators
  • Enzyme Inhibitors
  • NF-E2-Related Factor 2
  • Nfe2l2 protein, rat
  • Protoporphyrins
  • RNA, Messenger
  • zinc protoporphyrin
  • cobaltiprotoporphyrin
  • Heme Oxygenase (Decyclizing)
  • Hmox1 protein, rat