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目的研究铅硒共同孵育对海马神经元存活率、轴突长度及神经细胞粘附分子(NCAM)蛋白表达的影响。方法取出生24 h Wistar乳鼠,建立原代培养的海马神经元,通过氯化铅和硒蛋氨酸染毒后,用四甲基偶氮唑盐比色法测定神经元存活率,用Image pro-plus软件测定突起长度,并用蛋白质印迹法测定NCAM蛋白表达量。结果在培养第2 d,铅硒共同孵育组中神经元轴突长度比氯化铅组增长约6～17μm,差异有统计学意义(P<0.05);随着硒蛋氨酸剂量的增加,NCAM表达量也随之增加,在次高剂量组达最高(蛋白印迹的平均灰度比值为2.00),较氯化铅组增加了23.82%,然后依次为中剂量组(1.96)、低剂量组(1.89)、次低剂量组(1.80)和高剂量组(1.75),且NCAM140表达量明显高于NCAM180。结论硒可促进轴突的向外生长,并增加海马神经元NCAM的表达。
Objective To investigate the effects of co-incubation of lead and selenium on the survival rate of neurons, the length of axons and the expression of neural cell adhesion molecule (NCAM) in hippocampus. Methods Twenty-four-day-old Wistar rats were used to establish primary cultured hippocampal neurons. After exposure to lead chloride and selenomethionine, the survival rate of neurons was determined by MTT colorimetric assay. plus software to measure protrusion length and protein expression of NCAM by Western blotting. Results On day 2 of culture, the neuronal axon length in lead-selenium co-incubation group increased by 6-17 μm compared with lead chloride group (P <0.05). With the increase of selenomethionine dose, NCAM expression (The average gray value of the Western blot was 2.00), which was 23.82% higher than that of the lead chloride group, followed by the middle dose group (1.96) and the low dose group (1.89) ), Low dose group (1.80) and high dose group (1.75), and the expression of NCAM140 was significantly higher than NCAM180. Conclusion Selenium can promote axon outgrowth and increase the expression of NCAM in hippocampal neurons.