Heroin activates ATF3 and CytC via c-Jun N-terminal kinase pathways to mediate neuronal apoptosis

Med Sci Monit Basic Res. 2015 Apr 7;21:53-62. doi: 10.12659/MSMBR.893827.

Abstract

BACKGROUND Drug abuse and addiction has become a major public health problem that impacts all societies. The use of heroin may cause spongiform leukoencephalopathy (SLE). MATERIAL AND METHODS Cerebellar granule cells were derived from 7-day-old Sprague-Dawley rat pups. Neurons were dissociated from freshly dissected cerebella by mechanical disruption in the presence of 0.125% trypsin and DNaseI and then seeded at a density of 4×10^6 cells/ml in Dulbecco's modified Eagle's medium/nutrient mixture F-12 ham's containing 10% fetal bovine serum and Arc-C(sigma) at concentrations to inhibit glial cell growth inoculated into 6-well plates and a small dish. RESULTS We found that heroin induces the apoptosis of primary cultured cerebellar granule cells (CGCS) and that the c-Jun N-terminal kinase (JNK) pathway was activated under heroin treatment and stimulated obvious increases in the levels of C-jun, Cytc, and ATF3mRNA. CYTC and ATF3 were identified as candidate targets of the JNK/c-Jun pathway in this process because the specificity inhibitors SP600125 of JNK/C-jun pathways reduced the levels of C-jun, Cytc, and ATF3mRNA. The results suggested that SP600125 of JNK/C-jun can inhibit heroin-induced apoptosis of neurons. CONCLUSIONS The present study analyzes our understanding of the critical role of the JNK pathway in the process of neuronal apoptosis induced by heroin, and suggests a new and effective strategy to treat SLE.

MeSH terms

  • Activating Transcription Factor 3 / metabolism*
  • Animals
  • Apoptosis / drug effects*
  • Cytochromes c / metabolism*
  • DNA Primers / genetics
  • Fluorescent Antibody Technique
  • Heroin / pharmacology*
  • In Vitro Techniques
  • JNK Mitogen-Activated Protein Kinases / metabolism*
  • Neurons / drug effects*
  • Neurons / pathology
  • Rats
  • Rats, Sprague-Dawley
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction / drug effects*

Substances

  • Activating Transcription Factor 3
  • Atf3 protein, rat
  • DNA Primers
  • Heroin
  • Cytochromes c
  • JNK Mitogen-Activated Protein Kinases