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

Pu H, Wang X, Su L, Ma C, Zhang Y, Zhang L, Chen X, Li X, Wang H, Liu X, Zhang J - Med Sci Monit Basic Res (2015)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Science and Research Education Center, First Affiliated Hospital of Xinjiang Medical University, Urumqi, Sinciang, China (mainland).

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.

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Related in: MedlinePlus

In vitro culture cerebellar granule cells after five days, By laser confocal detection and identification Neuronal cell (×400), DAPI nuclear staining is blue (A), β-tubulin III Cytoplasmic staining is green (B); A+B was condition of merge (C).
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f1-medscimonitbasicres-21-53: In vitro culture cerebellar granule cells after five days, By laser confocal detection and identification Neuronal cell (×400), DAPI nuclear staining is blue (A), β-tubulin III Cytoplasmic staining is green (B); A+B was condition of merge (C).

Mentions: On Day 5, DIV5 cells were examined and identified by laser confocal: β-tubulin expressed in cerebellar granule cell cytoplasm (CGCs), blue showed DAPI staining nuclei that can excite the nucleus (Figure 1A), the second antibody with fluorescence-excited cytoplasm was green (Figure 1B). Figure 1C shows synthesis results for the morphological structure of integral CGCs. The purity of cultured cerebellar granule cells was more than 90% and can be used for further experiments.


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

Pu H, Wang X, Su L, Ma C, Zhang Y, Zhang L, Chen X, Li X, Wang H, Liu X, Zhang J - Med Sci Monit Basic Res (2015)

In vitro culture cerebellar granule cells after five days, By laser confocal detection and identification Neuronal cell (×400), DAPI nuclear staining is blue (A), β-tubulin III Cytoplasmic staining is green (B); A+B was condition of merge (C).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4400970&req=5

f1-medscimonitbasicres-21-53: In vitro culture cerebellar granule cells after five days, By laser confocal detection and identification Neuronal cell (×400), DAPI nuclear staining is blue (A), β-tubulin III Cytoplasmic staining is green (B); A+B was condition of merge (C).
Mentions: On Day 5, DIV5 cells were examined and identified by laser confocal: β-tubulin expressed in cerebellar granule cell cytoplasm (CGCs), blue showed DAPI staining nuclei that can excite the nucleus (Figure 1A), the second antibody with fluorescence-excited cytoplasm was green (Figure 1B). Figure 1C shows synthesis results for the morphological structure of integral CGCs. The purity of cultured cerebellar granule cells was more than 90% and can be used for further experiments.

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Science and Research Education Center, First Affiliated Hospital of Xinjiang Medical University, Urumqi, Sinciang, China (mainland).

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.

Show MeSH
Related in: MedlinePlus