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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 ,与细胞内胆红素含量不同有关
Objective To observe the toxic effects and mechanism of bilirubin on different types of neurons in rats. Methods Primary cultured rat cerebellar granule neurons, cortical neurons and hippocampal neurons were cultured in vitro. The effects of low concentration (0 ~ 17.1 μmol / L) bilirubin on the neurotoxicity were observed. The survival rate of neurons was measured by fluorescein diacetate staining, and the content of bilirubin was measured by fluorescence spectrophotometry. Results Bilirubin induced the death of cerebellar granule neurons in a dose-dependent manner. The survival rates of granule neurons were (100 ± 3.1)% at bilirubin concentrations of 4.3, 8.6 and 17.1 μmol / L, respectively , (18.5 ± 2.3)% and (2.0 ± 2.4%), respectively. The survival rates of cortical neurons were (90.7 ± 7.8)%, (85.3 ± 5.3)% and 74.5 ± 2 .4%). The survival rates of hippocampal neurons were (75 ± 6)%, (62 ± 5)% and (49 ± 5)%, respectively. Bilirubin on cerebral cortex neurons and hippocampal neurons was significantly lower than the neurons. After adding bilirubin for 3h, the fluorescence intensity of bilirubin in granule neurons, cortex and hippocampal neurons were 33.4 ± 2 .3, 10.7 ± 3.0, and 2.19 ± 4.3, respectively. The bilirubin in cerebellar granule neurons Superoxide dismutase was significantly higher than hippocampus and cortical neurons (P <0.01). Conclusions Cerebellar granule neurons are more sensitive to bilirubin than hippocampus and cortical neurons, and have different contents of bilirubin