PD150606 protects against ischemia/reperfusion injury by preventing μ-calpain-induced mitochondrial apoptosis

Arch Biochem Biophys. 2015 Nov 15:586:1-9. doi: 10.1016/j.abb.2015.06.005. Epub 2015 Jun 16.

Abstract

Calpain plays an important role in myocardial ischemia/reperfusion (I/R) injury. PD150606, a nonpeptide, cell-permeable and noncompetitive calpain inhibitor, has been shown to have protective properties in ischemic disease. The aims of the present study were to investigate whether PD150606 could alleviate myocardial I/R injury and to examine the possible mechanisms involved. The I/R model was established in vivo in C57BL/6 mice and in vitro using neonatal mouse cardiomyocytes, respectively. To evaluate the protective effects of PD150606 on I/R injury, we measured the myocardial infarct area, apoptosis, and expression of cleaved caspase-3. We also investigated the underlying mechanisms by examining mitochondrial function as reflected by the ATP concentration, translocation of cytochrome c, dynamics of mPTP opening, and membrane potential (ΔΨm), coupled with calpain activity. Pretreatment with PD150606 significantly reduced the infarct area and apoptosis caused by I/R. PD150606 pretreatment also reduced mitochondrial dysfunction by inhibiting calpain activation. Moreover, we found that μ-calpain is the main contributor to I/R-induced calpain activation. Knockdown of μ-calpain with siRNA significantly reversed calpain activation, mitochondrial dysfunction, and cardiomyocyte apoptosis caused by I/R in vitro. Our results suggest that PD150606 may protect against I/R injury via preventing μ-calpain-induced mitochondrial apoptosis.

Keywords: Apoptosis; Ischemia/reperfusion; Mitochondrial dysfunction; PD150606; μ-Calpain.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acrylates / pharmacology*
  • Animals
  • Animals, Newborn
  • Apoptosis / drug effects
  • Calpain / antagonists & inhibitors*
  • Calpain / genetics
  • Cardiotonic Agents / pharmacology*
  • Cell Hypoxia / drug effects
  • Cysteine Proteinase Inhibitors / pharmacology
  • Disease Models, Animal
  • Gene Knockdown Techniques
  • In Vitro Techniques
  • Male
  • Membrane Potential, Mitochondrial / drug effects
  • Mice
  • Mice, Inbred C57BL
  • Mitochondria, Heart / drug effects
  • Mitochondria, Heart / enzymology
  • Mitochondrial Membrane Transport Proteins / drug effects
  • Mitochondrial Permeability Transition Pore
  • Myocardial Reperfusion Injury / enzymology
  • Myocardial Reperfusion Injury / pathology
  • Myocardial Reperfusion Injury / prevention & control*
  • Myocytes, Cardiac / drug effects
  • Myocytes, Cardiac / enzymology
  • Myocytes, Cardiac / pathology
  • RNA, Small Interfering / genetics

Substances

  • Acrylates
  • Cardiotonic Agents
  • Cysteine Proteinase Inhibitors
  • Mitochondrial Membrane Transport Proteins
  • Mitochondrial Permeability Transition Pore
  • PD 150606
  • RNA, Small Interfering
  • Calpain
  • mu-calpain