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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

Crebellar granule cells apoptosis when maintained in different of heroin. DIV7 CGNs were then switched to medium containing 10 ug/ml (B), 40 ug/ml (C), 80 ug/ml (D), 120 ug/ml (E) heroin. Flow cytometry show neurons apoptosis maintained in different of heroin dosage for 24h. Control cells (A) were maintained for 24 h in heroin-free medium. The percentages of neurons apoptosis under the treatments indicated were quantified (F). Scale bar=10 um. Data represent the means ±SEM of three independent experiments. * p<0.05,** p<0.01.
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f3-medscimonitbasicres-21-53: Crebellar granule cells apoptosis when maintained in different of heroin. DIV7 CGNs were then switched to medium containing 10 ug/ml (B), 40 ug/ml (C), 80 ug/ml (D), 120 ug/ml (E) heroin. Flow cytometry show neurons apoptosis maintained in different of heroin dosage for 24h. Control cells (A) were maintained for 24 h in heroin-free medium. The percentages of neurons apoptosis under the treatments indicated were quantified (F). Scale bar=10 um. Data represent the means ±SEM of three independent experiments. * p<0.05,** p<0.01.

Mentions: To define experimental conditions of moderate neurotoxicity, we performed a dose-response analysis of cell viability following flow cytometry: After 24 h, neuronal cells remained normal in the no-heroin medium (Figure 3A). Apoptosis rates reached 5.23% with 10 ug/ml heroin concentration (Figure 3B) and 11.5% in 40 ug/ml heroin medium (Figure 3C). However, the average apoptosis rates were 46.3% and 57% in 80 ug/mL and 120 ug/mL heroin concentrations, respectively (Figure 3D, 3E), and the model groups were significantly different compared with the control group (** P<0.01). We proved that these cells were undergoing apoptosis. With these criteria, the results consistently demonstrated that heroin in our model system triggered CGC apoptosis. Heroin-induced CGC apoptosis rates were almost the same as morphological changes, which indicated that the apoptosis rates rapidly increased when the concentration of heroin increased.


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)

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

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

f3-medscimonitbasicres-21-53: Crebellar granule cells apoptosis when maintained in different of heroin. DIV7 CGNs were then switched to medium containing 10 ug/ml (B), 40 ug/ml (C), 80 ug/ml (D), 120 ug/ml (E) heroin. Flow cytometry show neurons apoptosis maintained in different of heroin dosage for 24h. Control cells (A) were maintained for 24 h in heroin-free medium. The percentages of neurons apoptosis under the treatments indicated were quantified (F). Scale bar=10 um. Data represent the means ±SEM of three independent experiments. * p<0.05,** p<0.01.
Mentions: To define experimental conditions of moderate neurotoxicity, we performed a dose-response analysis of cell viability following flow cytometry: After 24 h, neuronal cells remained normal in the no-heroin medium (Figure 3A). Apoptosis rates reached 5.23% with 10 ug/ml heroin concentration (Figure 3B) and 11.5% in 40 ug/ml heroin medium (Figure 3C). However, the average apoptosis rates were 46.3% and 57% in 80 ug/mL and 120 ug/mL heroin concentrations, respectively (Figure 3D, 3E), and the model groups were significantly different compared with the control group (** P<0.01). We proved that these cells were undergoing apoptosis. With these criteria, the results consistently demonstrated that heroin in our model system triggered CGC apoptosis. Heroin-induced CGC apoptosis rates were almost the same as morphological changes, which indicated that the apoptosis rates rapidly increased when the concentration of heroin increased.

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