Pu Hongwei, Wang Xuemei, Su Liping, Ma Chuang, Zhang Yan, Zhang Liping, Chen Xiao, Li Xiujuan, Wang Hua, Liu Xiaoshan, Zhang Jianlong
Department of Science and Research Education Center, First Affiliated Hospital of Xinjiang Medical University, Urumqi, Sinciang, China (mainland).
Xinjiang Key Laboratory of Medical Animal Model Research, First Affiliated Hospital of Xinjiang Medical University, Urumqi, Sinciang, China (mainland).
Med Sci Monit Basic Res. 2015 Apr 7;21:53-62. doi: 10.12659/MSMBR.893827.
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.
背景 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。海洛因的使用可能会导致海绵状白质脑病(SLE)。
材料与方法 小脑颗粒细胞取自7日龄的Sprague-Dawley大鼠幼崽。在0.125%胰蛋白酶和脱氧核糖核酸酶I存在的情况下,通过机械破碎从刚解剖的小脑中分离出神经元,然后以4×10^6个细胞/毫升的密度接种于含有10%胎牛血清和用于抑制胶质细胞生长的Arc-C(西格玛)的杜尔贝科改良伊格尔培养基/营养混合物F-12哈姆氏培养基中,接种到6孔板和一个小培养皿中。
结果 我们发现海洛因可诱导原代培养的小脑颗粒细胞(CGCS)凋亡,并且在海洛因处理下c-Jun氨基末端激酶(JNK)通路被激活,刺激C-jun、细胞色素C(Cytc)和活化转录因子3(ATF3)mRNA水平显著升高。在此过程中,CYTC和ATF3被确定为JNK/c-Jun通路的候选靶点,因为JNK/C-jun通路的特异性抑制剂SP600125降低了C-jun、Cytc和ATF3mRNA的水平。结果表明,JNK/C-jun的SP600125可抑制海洛因诱导的神经元凋亡。
结论 本研究分析了我们对JNK通路在海洛因诱导的神经元凋亡过程中关键作用的理解,并提出了一种治疗SLE的新的有效策略。