乙醇对体外培养大鼠大脑皮层神经细胞分化的影响  

The effect of ethanol on the differentiation of rat cortex neurons in vitro culture

作　　者：贾景娜[1] 史为博[1] 易善勇[1] 王冉[2] 张国忠[1] 马春玲[1] 左敏[1] 李英敏[1] 

机构地区：[1]河北医科大学法医学系,河北法医学重点实验室及法医鉴定,河北省法医分子鉴定协同创新中心,石家庄050017 [2]河北医科大学第一医院精神科,石家庄050031

出　　处：《神经解剖学杂志》2016年第3期345-351,共7页Chinese Journal of Neuroanatomy

基　　金：河北省自然科学基金(C2009001103;H2013206215)

摘　　要：目的:观察神经细胞微管相关蛋白2(microtubeasso ciated protein-2,MAP2)、孤儿核受体1(nuclear receptor-related factor 1,NURR1)、拓扑异构酶Ⅱβ(DNA topoisomereⅡβ,TopoⅡβ)在乙醇毒性作用下的表达变化,探讨MAP2、NURR1、TopoⅡβ与乙醇导致的神经细胞分化异常的相关关系。方法:取新生鼠(出生24 h内),75%酒精消毒,断头取脑,分离皮层神经细胞接种于培养板,给予剂量75 mmol/L的乙醇培养3 d、5 d、7 d后,采用免疫荧光观察MAP2、NURR1、TopoⅡβ的表达变化;Fluoro-Jade B荧光染色观察神经细胞变性坏死情况。结果:随乙醇染毒时间的延长,原代培养的神经细胞分布较正常对照组稀疏,MAP2的表达发生了变化,以7 d组变化最明显,突起变细变短,阳性表达减弱;NURR1及TopoⅡβ的表达也随乙醇染毒时间的延长,阳性细胞数目及阳性信号表达的强度均呈下降趋势,以7 d最为显著。NURR1阳性细胞计数为(9.6±3.5)与正常对照组的(45.2±3.96)有显著差异(P<0.05)。TopoⅡβ(55.8±3.77)与正常对照组的(86.2±4.82)有明显差异(P<0.05);Fluoro-Jade B荧光标记显示在乙醇染毒5 d组及7 d组均可见阳性细胞。结论:MAP2、NURR1、TopoⅡβ可能参与了乙醇导致的神经细胞分化异常,改变了细胞骨架稳定,最终导致了部分神经细胞的变性坏死。Objective： To observe the changes of microtubeasso ciated protein-2（ MAP2）,nuclear receptor-related factor 1（ NURR1） and DNA topoisomereⅡβ（ TopoⅡβ） under the toxicity of ethanol,and finally explore the correlation of MAP2,NURR1,TopoⅡβ and abnormal nerve cell differentiation induced by ethanol. Methods： The newborn rats（ born within 24 h） were disinfected with 75%（ V/V） ethanol and decapitated. Isolated cortex neurons were planted on culture plates and treated with 75 mmol / L ethanol for 3 d,5 d and 7 d. Immunofluorescence was used to observe the changes of MAP2,NURR1 and TopoⅡβ. Fluoro-Jade B staining was used to detect neuronal degeneration and necrosis.Results： With prolonged ethanol exposure,the distribution of nerve cells was sparse compared with control group,and the expression of MAP2 was decreased. Neuronal processes were thinner and shorter,which was the most obvious in 7 d group. Compared with the control group（ 45. 2 ± 3. 96）,the number of NURR1 positive cells（ 9. 6 ± 3. 5） was signifi-cantly decreased（ P〈0. 05）. Besides,the number of TopoⅡβ positive cells（ 55. 8 ± 3. 77） was obviously difference（ P〈0. 05） compared with the control group（ 86. 2 ± 4. 82）. With Fluoro-Jade B fluorescence staining,the degeneration and necrosis of nerve cells were found in 5 d and 7 d group. Conclusion： MAP2,NURR1 and TopoⅡβ are associated with the abnormal differentiation of nerve cells induced by ethanol,which disordered the stability of the cytoskeleton,and eventually lead to the degeneration and necrosis of nerve cells.

关 键 词：原代培养 乙醇 神经细胞分化 FLUORO-JADE B荧光标记 大鼠 

分 类 号：R749.62[医药卫生—神经病学与精神病学]