氨对体外培养胚胎大鼠大脑皮质神经元凋亡的影响

目的通过观察体外培养的胚胎大鼠大脑皮质神经细胞在氨作用下的形态学、超微结构的变化,探讨氨与细胞凋亡及肝性脑病的关系.方法不同浓度的氨作用于体外培养的18d胎龄胚胎大鼠大脑皮质神经细胞24h后,采用光镜、透射电镜观察细胞形态及超微结构;利用流式细胞仪进行凋亡细胞定量观察.结果对照组与实验组在光镜下形态不同,两组在透射电镜下可见到典型的"凋亡小体”,流式细胞仪定量显示出两组凋亡细胞数量具有显著性差异(P＜0.01),且不同浓度的氨引起的神经细胞凋亡数量也存在显著性差异(P＜0.01).结论氨可诱导体外培养神经细胞产生过量的细胞凋亡.

Apoptosis of Cerebral Cortex Neuron Induced by Ammonia in Vitro in Embryonic Rat

Objective: Our aims were to observe the apoptosis in the neuron induced by ammonia in vitro and to explore the pathogenesis of hepatic encephalopathy. Methods: Cerebral cortex neurons from embryonic Wistar rat (18 days after birth) were cultured in vitro.After 3 days,cells were treated with ammonia for 24 hours. The morphology and ultrastructure of apoptosis cells were observed by microscope and transmission electron microscope.Flow cytometry was used to count apoptosis cells. Results: Neurons of controls grew well.Neurosomes were full and neucleus were round or oval. But in experimental group, neurons were significantly smaller even exfoliation.Crescent concentrational chromatin were located on nuclear membrane."Apoptotic bodies"were seen with electron microscope.The amount of apoptosis cells showed stastistic difference between control and experimental groups(P<0.01). The amounts of apoptosis cells induced by different concentration of ammonia were different(P<0.01). Conclusion: More neuronal apoptosis may be induced by ammonia in vitro.It may provide experimental basis for further research of pathogenesis of hepatic encephalopathy.