Ischemic preconditioning modulates the expression of several genes, leading to the overproduction of IL-1Ra, iNOS, and Bcl-2 in a human model of liver ischemia-reperfusion

FASEB J. 2005 Oct;19(12):1617-26. doi: 10.1096/fj.04-3445com.

Abstract

Ischemia triggers an inflammatory response that precipitates cell death during reperfusion. Several studies have shown that tissues are protected by ischemic preconditioning (IP) consisting of 10 min of ischemia followed by 10 min of reperfusion just before ischemia. The molecular basis of this protective effect is poorly understood. We used cDNA arrays (20K) to compare global gene expression in liver biopsies from living human liver donors who underwent IP (n=7) or not (n=7) just before liver devascularization. Microarray data were analyzed using pairedt test with a type I error rate fixed at alpha = 2.5 10(6) (Bonferroni correction). We found that 60 genes were differentially expressed (36 over- and 24 underexpressed in preconditioning group). After IP, the most significantly overexpressed gene was IL-1Ra. This was confirmed by immunoblotting. Differentially expressed were genes involved in apoptosis (NOD2, ephrin-A1, and calpain) and in the carbohydrate metabolism. A significant increase in the amount of the anti-apoptotic protein Bcl-2 in preconditioned livers but no change in the cleavage of procaspase-3, -8, and -9 was observed. We also observed an increase in the amount in the inducible nitric oxide synthase. Therefore, the benefits of IP may be associated with the overproduction of IL-1Ra, Bcl-2, and NO countering the proinflammatory and proapoptotic effects generated during ischemia-reperfusion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Apoptosis
  • Biopsy
  • Blotting, Western
  • Carbohydrates / chemistry
  • Caspase 3
  • Caspase 8
  • Caspase 9
  • Caspases / metabolism
  • DNA, Complementary / metabolism
  • Ephrin-A1 / metabolism
  • Female
  • Gene Expression Regulation*
  • Humans
  • Immunoblotting
  • Immunohistochemistry
  • Inflammation
  • Interleukin 1 Receptor Antagonist Protein
  • Ischemic Preconditioning*
  • Liver / metabolism
  • Liver / pathology*
  • Male
  • Middle Aged
  • Nitric Oxide / metabolism
  • Nitric Oxide Synthase / metabolism
  • Nitric Oxide Synthase Type II / metabolism*
  • Oligonucleotide Array Sequence Analysis
  • Proto-Oncogene Proteins c-bcl-2 / metabolism*
  • RNA, Messenger / metabolism
  • Reperfusion
  • Reperfusion Injury*
  • Sialoglycoproteins / metabolism*
  • Time Factors

Substances

  • Carbohydrates
  • DNA, Complementary
  • Ephrin-A1
  • IL1RN protein, human
  • Interleukin 1 Receptor Antagonist Protein
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Messenger
  • Sialoglycoproteins
  • Nitric Oxide
  • Nitric Oxide Synthase
  • Nitric Oxide Synthase Type II
  • CASP3 protein, human
  • CASP8 protein, human
  • CASP9 protein, human
  • Caspase 3
  • Caspase 8
  • Caspase 9
  • Caspases