摘要
目的 研究无镁处理诱导反复惊厥样放电对体外培养的发育中神经元造成的损伤。方法 培养的胚胎鼠大脑皮层神经元为研究对象 ,应用锥虫蓝染色、乳酸脱氢酶 (LDH)活浓度测定、流式细胞计数、噻唑蓝 (MTT)代谢率测定 ,观察神经元坏死、凋亡以及功能损伤。结果 ( 1)体外培养不同阶段 ( 6、12、17d)神经元无镁处理后不同时间 ( 6、2 4、72h) ,存活神经元数目较对照组略减少 ,LDH活浓度较对照组略升高 ,但差异均无显著性 (P均 >0 0 5 )。 ( 2 )体外培养不同阶段无镁处理后各时间点神经元凋亡较对照组略增加 ,但差异无显著性 (P >0 0 5 )。 ( 3 )体外培养 6d ,无镁处理后 6h ,MTT代谢率较对照组降低 (P <0 0 5 ) ,为对照组的 86 4% ;体外培养 12、17d ,于无镁处理后 2 4h ,较对照组降低 (P <0 0 5 ) ,分别为对照组的 78 7%、70 9%。结论 无镁处理诱导的反复惊厥样放电引起的发育中神经元损伤主要是功能损伤。发育早期的神经元损伤发生早 ,但程度轻 ;发育相对成熟的神经元损伤发生晚 。
Objective To study cortical neuron injury following recurrent epileptiform discharges induced by magnesium-free treatment in vitro. Methods Cultured embryo cortical neurons were exposed to magnesium-free media for 3 h, then they were returned to regular media containing normal level magnesium. At different time after Mg 2+-free treatment, trypan blue staining and determination of LDH activity were used to determine the cell viability,flow cytometry was applied to measure neuronal apoptosis, and MTT assay to study metabolic rate. Results (1)Neuronal morphology on light microscopy following Mg 2+-free treatment showed that there were no prominent alterations. (2) At different time (6, 12, 72 h) after Mg 2+-free treatment, neuronal viability by trypan blue staining and LDH activity showed modest changes compared with time-matched control in different culture days (6, 12, 17 d) (P>0.05). (3) Cell apoptosis increased mildly at different time after Mg 2+-free treatment in neurons cultured for different days, but the increase was not significant (P>0.05). (4) Metabolic rate decreased at 6 h after Mg 2+-free treatment (P<0.05) in neurons cultured for 6 d, and was 86.4% of that of the control; while the rate at 24 h in neurons cultured for 12 d and 17 d also decreased (P<0.05), being 78.7% and 70.9%, respectively, of that of the control. Conclusions These findings demonstrated that the injury occurred on cultured cortical neurons caused by magnesium-free-treatment-induced recurrent epileptiform discharges was mainly functional and relatively mature neurons displayed more severe and much later mitochondrial function impairment than immature neurons.
出处
《中华儿科杂志》
CAS
CSCD
北大核心
2003年第1期25-27,共3页
Chinese Journal of Pediatrics
