亚低温对缺血性神经元凋亡后凋亡诱导因子表达的影响  

Effect of mild hypothermia on apoptosis-inducing factor in ischemic neuronal apoptosis in vitro

作　　者：张艳[1] 王德生[1] 汤颖[1] 

机构地区：[1]哈尔滨医科大学第一临床医学院神经内科,黑龙江哈尔滨150001

出　　处：《中风与神经疾病杂志》2007年第5期534-536,641,共4页Journal of Apoplexy and Nervous Diseases

基　　金：黑龙江省博士后基金资助(LBH-Z05165)

摘　　要：目的研究亚低温对凋亡诱导因子(AIF)介导的缺血性神经元凋亡通路的影响。方法建立离体大鼠皮质神经元凋亡模型。皮质神经元原代培养,分为常温组(37℃)和亚低温组(33℃),在剥夺血清后的8h、16h和24h,测定神经元生存率;应用免疫细胞化学的方法检测微管相关蛋白-2(MAP-2);通过Western Blotting方法测定神经元的细胞核和细胞浆中AIF的含量变化。结果缺血24h组,神经元生存率分别为42.34%(37℃)和65.76% (33℃),差异有统计学意义(P<0.05)。随着缺血时间的延长,MAP-2的表达逐渐减低,在缺血16h和24h,两组间差异有统计学意义(P<0.05)。细胞核中AIF的含量逐渐增加,而细胞浆中AIF的含量逐渐减少,在缺血16h和24h,与常温组相比,低温组细胞核AIF的含量显著性减低(P<0.05)。结论亚低温能促进缺血后神经元的存活,减少MAP-2的降解。亚低温干预了AIF介导的细胞凋亡途径,减少了缺血性损伤导致的AIF核位移。Objective The aim of this study was to investigate the effects of post-ischemic hypothermia on apoptotic neuronal cell death,microtubule-associated protein-2（MAP-2） and expression of apoptosis-inducing protein （AIF） after neuronal ischemia in vitro. Methods The model of cortical neuronal apoptosis was prepared by serum deprivation of neuronal culture of rat. The neurons were kept at 37℃ or 33℃ for 24 hours. The neuron viability was determined by MTT test. The immunoreactivity of MAP-2 was examined by immunocytochemistry. The detection of AIF nuclear translocation was by Western Blot. Results The cell viability was respectively 42.34% and 65. 76% at 33℃ and 37℃ after 24 hours,and there was significant difference between the two groups（P〈 0. 05）. The areas of MAP-2 immunostaining decreased gradually in the two groups ,but the change was more obvious at 37℃. There was significant difference between the two groups（P〈0.05） after 16 and 24 hours. The immunoreactivity of cytosolic AIF decreased gradually by Western Blot,and was at the lowest level at 24h. There was significant difference in nuclear and cytosolic AIF between the two groups（P〈0. 05） after 16 and 24 hours. Conclusion Mild hypothermia may improve neuron viability,and prevent neuronal apoptosis. Mild hypothermia ean reduee MAP-2 loss,and keep neuron morphology normal. AIF may be involved in neuronal cell death after ischemia and AIF nuclear translocation can be inhibited by mild hypothermia. Mild hypothermia may act through the inhibition of the caspase-independent pathway.

关 键 词：亚低温 神经元凋亡 凋亡诱导因子(AIF) 

分 类 号：R743.31[医药卫生—神经病学与精神病学]