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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

Cerebellar granule cells die when maintained in different of heroin (×200). DIV7 CGNs were then switched to medium containing 10 ug/ml, 40 ug/ml 80 ug/ml, 120 ug/ml heroin. Phase-contrast micrographs show neurons maintained in 10 ug/ml (B), 40 ug/ml (C), 80 ug/ml (D), 120 ug/ml (E) heroin for 24. Control cells (A) were maintained for 24 h in heroin-free medium. The percentages of death neurons under the treatments indicated were quantified (F). Scale bar=10 um. Data represent the means ±SEM of five independent experiments. * p<0.05,** p<0.01.
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f2-medscimonitbasicres-21-53: Cerebellar granule cells die when maintained in different of heroin (×200). DIV7 CGNs were then switched to medium containing 10 ug/ml, 40 ug/ml 80 ug/ml, 120 ug/ml heroin. Phase-contrast micrographs show neurons maintained in 10 ug/ml (B), 40 ug/ml (C), 80 ug/ml (D), 120 ug/ml (E) heroin for 24. Control cells (A) were maintained for 24 h in heroin-free medium. The percentages of death neurons under the treatments indicated were quantified (F). Scale bar=10 um. Data represent the means ±SEM of five independent experiments. * p<0.05,** p<0.01.

Mentions: To investigate the effect of heroin, CGCs were cultured for 7 days in vitro. The mature neurons were then shifted to media containing 10 ug/ml (b), 40 ug/ml (c), 80 ug/ml (d), and 120 ug/ml (e) heroin for 24 h. In the medium without heroin, the neurons were healthy (Figure 2A). Within 24 h of switching to 10 ug/ml heroin-containing media, neurons underwent apoptosis and the apoptotic rate was higher than in the control group (Figure 2B). In 40-ug/ml heroin media, cell volume was shrunken (Figure 2C). With a dose of 80 ug/ml heroin, most of the CGCs were undergoing apoptosis (Figure 2D). However, more than 50% of cells were shrunken in 120 ug/ml heroin media with the disappearances of grid structures (Figure 2E). The morphological changes accompanying death of granule neurons are the characteristics of cells undergoing apoptosis.


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)

Cerebellar granule cells die when maintained in different of heroin (×200). DIV7 CGNs were then switched to medium containing 10 ug/ml, 40 ug/ml 80 ug/ml, 120 ug/ml heroin. Phase-contrast micrographs show neurons maintained in 10 ug/ml (B), 40 ug/ml (C), 80 ug/ml (D), 120 ug/ml (E) heroin for 24. Control cells (A) were maintained for 24 h in heroin-free medium. The percentages of death neurons under the treatments indicated were quantified (F). Scale bar=10 um. Data represent the means ±SEM of five independent experiments. * p<0.05,** p<0.01.
© Copyright Policy
Related In: Results  -  Collection

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

f2-medscimonitbasicres-21-53: Cerebellar granule cells die when maintained in different of heroin (×200). DIV7 CGNs were then switched to medium containing 10 ug/ml, 40 ug/ml 80 ug/ml, 120 ug/ml heroin. Phase-contrast micrographs show neurons maintained in 10 ug/ml (B), 40 ug/ml (C), 80 ug/ml (D), 120 ug/ml (E) heroin for 24. Control cells (A) were maintained for 24 h in heroin-free medium. The percentages of death neurons under the treatments indicated were quantified (F). Scale bar=10 um. Data represent the means ±SEM of five independent experiments. * p<0.05,** p<0.01.
Mentions: To investigate the effect of heroin, CGCs were cultured for 7 days in vitro. The mature neurons were then shifted to media containing 10 ug/ml (b), 40 ug/ml (c), 80 ug/ml (d), and 120 ug/ml (e) heroin for 24 h. In the medium without heroin, the neurons were healthy (Figure 2A). Within 24 h of switching to 10 ug/ml heroin-containing media, neurons underwent apoptosis and the apoptotic rate was higher than in the control group (Figure 2B). In 40-ug/ml heroin media, cell volume was shrunken (Figure 2C). With a dose of 80 ug/ml heroin, most of the CGCs were undergoing apoptosis (Figure 2D). However, more than 50% of cells were shrunken in 120 ug/ml heroin media with the disappearances of grid structures (Figure 2E). The morphological changes accompanying death of granule neurons are the characteristics of cells undergoing apoptosis.

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