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07 April 2015 : Original article  

Heroin Activates ATF3 and CytC via c-Jun N-Terminal Kinase Pathways to Mediate Neuronal Apoptosis

Hongwei PuABCD, Xuemei WangADF, Liping SuD, Chuang MaB, Yan ZhangD, Liping ZhangE, Xiao ChenF, Xiujuan LiB, Hua WangB, Xiaoshan LiuAF, Jianlong ZhangAF

DOI: 10.12659/MSMBR.893827

Med Sci Monit Basic Res 2015; 21:53-62

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.

Keywords: Activating Transcription Factor 3 - metabolism, Cytochromes c - metabolism, DNA Primers - genetics, Fluorescent Antibody Technique, Heroin - pharmacology, In Vitro Techniques, JNK Mitogen-Activated Protein Kinases - metabolism, Neurons - pathology, Real-Time Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction, Signal Transduction - drug effects

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Medical Science Monitor Basic Research eISSN: 2325-4416
Medical Science Monitor Basic Research eISSN: 2325-4416