Targeting intestinal epithelial cell-programmed necrosis alleviates tissue injury after intestinal ischemia/reperfusion in rats

J Surg Res. 2018 May:225:108-117. doi: 10.1016/j.jss.2018.01.007. Epub 2018 Feb 21.

Abstract

Background: Intestinal dysfunction, especially acute pathologies linked to intestinal ischemia/reperfusion (I/R) injury, is profoundly affected by inflammation and improper execution of cell death. Few studies have examined the efficacy of combined strategies in regulated intestinal epithelial necrosis after intestinal I/R. Here, we evaluated the functional interaction between poly (adenosine diphosphate-ribose) polymerase 1 (PARP-1)-induced parthanatos and receptor-interacting protein 1/3 (RIP1/3) kinase-induced necroptosis in the pathophysiological course of acute ischemic intestinal injury.

Methods: Anesthetized adult male Sprague-Dawley rats were subjected to superior mesenteric artery occlusion consisting of 1.5 h of ischemia and 6 h of reperfusion. The PARP-1-specific inhibitor PJ34 (10 mg/kg) and the RIP1-specific inhibitor Necrostatin-1 (1 mg/kg) were intraperitoneally administered 30 min before the induction of ischemia.

Results: Intestinal I/R was found to result in PARP-1 activation and RIP1/3-mediated necrosome formation. PJ34 or Necrostatin-1 treatment significantly improved the mucosal injury, while the combined inhibition of PARP-1 and RIP1/3 conferred optimal protection of the intestine. Meanwhile, results from terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling assay showed a decrease in intestinal epithelial cell death. Interestingly, we further showed that PARP-1 might act as a downstream signaling molecule of RIP1 in the process of I/R-induced intestinal injury and that the RIP1/PARP-1-dependent cell death signaling pathway functioned independently of caspase 3 inhibition.

Conclusions: The results of our study provide a molecular basis for combination therapy that targets both pathways of regulated necrosis (parthanatos and necroptosis), to treat acute intestinal I/R-induced intestinal epithelial barrier disruption.

Keywords: Intestine; Ischemia/reperfusion injury; Necroptosis; Parthanatos; Regulated necrosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Disease Models, Animal
  • Drug Therapy, Combination / methods
  • Epithelial Cells / drug effects
  • Epithelial Cells / pathology*
  • Humans
  • Imidazoles / pharmacology*
  • Indoles / pharmacology*
  • Intestinal Mucosa / blood supply
  • Intestinal Mucosa / cytology
  • Intestinal Mucosa / drug effects
  • Intestinal Mucosa / pathology*
  • Male
  • Necrosis / drug therapy
  • Phenanthrenes / pharmacology*
  • Phenanthrenes / therapeutic use
  • Poly (ADP-Ribose) Polymerase-1 / antagonists & inhibitors
  • Poly (ADP-Ribose) Polymerase-1 / metabolism
  • Protein Serine-Threonine Kinases / antagonists & inhibitors
  • Protein Serine-Threonine Kinases / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Receptor-Interacting Protein Serine-Threonine Kinases / antagonists & inhibitors
  • Receptor-Interacting Protein Serine-Threonine Kinases / metabolism
  • Reperfusion Injury / drug therapy*
  • Reperfusion Injury / etiology
  • Reperfusion Injury / pathology
  • Signal Transduction / drug effects
  • Treatment Outcome

Substances

  • Imidazoles
  • Indoles
  • N-(oxo-5,6-dihydrophenanthridin-2-yl)-N,N-dimethylacetamide hydrochloride
  • Phenanthrenes
  • necrostatin-1
  • Parp1 protein, rat
  • Poly (ADP-Ribose) Polymerase-1
  • Protein Serine-Threonine Kinases
  • RIPK1 protein, rat
  • Receptor-Interacting Protein Serine-Threonine Kinases
  • Ripk3 protein, rat