胆红素对原代培养不同类型大鼠神经元的毒性作用  

The Toxic Effect of Bilirubin on Different Types of Neurons of Rat in Primary Culture

作　　者：李晓瑜[1] 林穗珍[1] 欧阳颖[1] 颜光美[2] 

机构地区：[1]中山医科大学附属第一医院儿科,广州510080 [2]中山医科大学附属第一医院药理教研室,广州510080

出　　处：《中华围产医学杂志》2002年第1期67-69,共3页Chinese Journal of Perinatal Medicine

基　　金：国家自然科学基金资助项目 (基金编号 39770 782 );广东省自然科学基金资助项目 (基金编号 970 0 5 2 )

摘　　要：目的　观察胆红素对大鼠不同类型神经元的毒性作用及机制。　方法　采用原代培养的大鼠小脑颗粒神经元、皮质神经元及海马神经元 ,观察低浓度 (0～ 17.1μm ol/ L)胆红素对其毒性的影响。用二乙酸荧光素染色法测定神经元存活率 ,荧光技术测定细胞内胆红素的含量。　结果　胆红素呈剂量依赖性地诱导小脑颗粒神经元死亡 ,在胆红素浓度为 4.3、8.6及 17.1μm ol/ L 时 ,颗粒神经元的存活率分别为 (5 0 .1± 3.1) %、(18.5± 2 .3) %和 (2 .0± 2 .4) % ;皮质神经元的存活率分别为(90 .7± 7.8) %、(85 .3± 5 .3) %和 (74.5± 2 .4) % ;海马神经元的存活率分别为 (75± 6 ) %、(6 2± 5 ) %和 (4 9± 5 ) %。胆红素对大脑皮质神经元及海马神经元的毒性明显低于颗粒神经元。加入胆红素 3h后 ,颗粒神经元、皮质及海马神经元细胞内胆红素的荧光强度分别为 33.4± 2 .3、10 .7± 3.0、2 1.9±4.3,小脑颗粒神经元内胆红素含量明显高于海马及皮质神经元 (P<0 .0 1)。　结论　小脑颗粒神经元较海马及皮质神经元对胆红素的毒性敏感 。Objective To investigate cytotoxicity and its mechanism of bilirubin on different types of neurons of rat in primary culture. Methods Toxic effect of bilirubin on primary cultured neurons (at cerebellar oranule hippocampal and cortical neurons) was observed. Ncuron survival was detected by fluorescei diacetate (FDA) staining. Measurement of fluorescent intensity of intracellular dilirudin were performed by microspectroflurimetry imaging system. Results Biliruin induced cytotoxicity to different type neurons in a dose-dependent manner. Exposed at 8d to different concentrations of bilirubin (4.3 μmol/L、8.6 μmol/L and 17.1 μmol/L); cerebellar gtanule neuron survival werc (50.1±3.1)%,(18.5±2.3)%, (2.0±2.4)%, cortical neuron survival were (90.7±7.8)%, (85.3±5.3)%,(74.5±2.4)%, and hippocampal neuron survival were (75±6)%, (62±5)%, (49±5)% respectively. The toxic effect of bilirubin on ccrebellar granule neuron was significantly higher than that on hippocampal and cortical neurons. The fluorescent intensity of bilirubin in gra- nule, hippocampal and cortical neurons were 33.4±2.3、10.7±3.0、21.9±4.3 respectively three hours after addition of bilirubin to the medium. Significantly higher bilirubin concentration was found in granule neurons than that in hippocampal and cortical neurons (P<0.01). Conclusion The cytotoxic effects may be associated with the content of intracellular bilirubin in neurons and this might contri bute to the apparent higher sensitivity to bilirubin cytotoxic effects in granule neurons than in hippocampal and cortical neurons.

关 键 词：胆红素 神经元 大鼠 毒性试验 原代培养 胆红素脑病 

分 类 号：R363[医药卫生—病理学]