Heroin activates ATF3 and CytC via c-Jun N-terminal kinase pathways to mediate neuronal apoptosis.
Heroin activates ATF3 and CytC via c-Jun N-terminal kinase pathways to mediate neuronal apoptosis.
复制标题
海洛因通过 c-Jun N 端激酶途径激活 ATF3 和 CytC 介导神经元凋亡
DOI：
10.12659/msmbr.893827
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发表时间：
2015-04-07
影响因子：
2.8
通讯作者：
Zhang J
中科院分区：
文献类型：
--
作者：
Pu H;Wang X;Su L;Ma C;Zhang Y;Zhang L;Chen X;Li X;Wang H;Liu X;Zhang J
药物滥用和成瘾已成为影响所有社会的一个主要公共卫生问题。海洛因的使用可引起海绵状白质脑病（SLE）。材料/方法小脑颗粒细胞来源于7日龄Sprague-Dawley大鼠幼仔。在0.125%胰蛋白酶和DNA酶I存在下，通过机械破碎从新鲜解剖的小脑中分离神经元，然后以4×106个细胞/ml的密度接种在含有10%胎牛血清和Arc-C（sigma）的Dulbecco改良Eagle培养基/营养混合物F-12 ham中，将其接种到6孔板和一个小培养皿中，浓度可抑制神经胶质细胞生长。结果海洛因可诱导原代培养的小脑颗粒细胞（CGCS）凋亡，并激活c-Jun N-末端激酶（JNK）通路，使CGCS细胞中C-jun、Cytc和ATF 3 mRNA水平明显升高。由于JNK/C-jun通路的特异性抑制剂SP 600125降低了C-jun、Cytc和ATF 3 mRNA的水平，因此在该过程中确定CYTC和ATF 3为JNK/c-Jun通路的候选靶点。提示JNK/C-jun的SP 600125对海洛因诱导的神经元凋亡有抑制作用。结论JNK信号通路在海洛因诱导的神经细胞凋亡过程中起重要作用，为治疗SLE提供了新的有效策略。
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/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 4×106 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.