摘要
目的 :探讨IL 1ra对体外发育不同阶段神经元惊厥性损伤的拮抗作用及可能机制。方法 :以培养的发育中皮层神经元为研究对象 ,MTT代谢率观察神经元损伤 ,激光共聚焦显微镜扫描检测神经元 [Ca2 + ]i,real time定量RT PCR方法测定NMDARmRNA表达。结果 :(1 )IL 1ra预处理后再给无镁 +IL 1ra处理 ,与同期单纯无镁组比较 ,6d的神经元MTT代谢率无明显改变 ,而 1 7d的神经元显著增高。 (2 )IL 1ra预处理后 ,6d、1 7d的神经元 [Ca2 + ]i峰值显著低于同期单纯无镁组 ,但拮抗 [Ca2 + ]i增高的作用在 6d的神经元较 1 7d的神经元更明显。 (3)IL 1ra预处理后 ,6d、1 7d的神经元NR1mRNA表达有所降低 ,且IL 1ra降低不同培养时间NR1mRNA的作用差异无显著性 ;用IL 1ra后 1 7d神经元的NR2AmRNA表达降低 ,与同期对照组比较差异无显著性 ,而 6d神经元NR2AmRNA表达仍高于同期对照组 ;IL 1ra对不同培养时间NR2BmRNA均无明显降低作用。结论 :IL 1ra对神经元惊厥性损伤的保护作用有一定的年龄依赖性。这种保护作用可能通过减轻细胞内Ca2 +
Objective: To study the protection of IL 1ra in cultured developing neuron injury following Mg 2+ free induced epileptiform discharges. Methods: Rat embryo cortical neurons cultured for 6 d and 17 d were directly exposed to Mg 2+ free media, or pretreated with IL 1 receptor antagonist or NMDA receptor antagonists before being exposed to Mg 2+ free media, and then returned to regular media.MTT assay was used to study mitochondrial function injury, laser scanning confocal microscope to measure [Ca 2+ ]i, and real time RT PCR to detect gene mRNA expression. Results:(1) MTT conversion rates were higher in neurons pre and co treated with 10 mg/L IL 1ra than those of neurons with only Mg 2+ free treatment in neurons cultured for 17 d, but not in neurons cultured for 6 d.(2) [Ca 2+ ]i was lower in neurons pre and co treated with 10 mg/L IL 1ra than those of neurons with only Mg 2+ free treatment, either in neurons cultured for 6 d or in neurons cultured for 17 d, and the effects of IL 1ra on [Ca 2+ ]i change were different between neurons cultured for 6 d and neurons cultured for 17 d.(3) Pre and co treated with 10 mg/L IL 1ra NR1 mRNA expression increase induced by Mg 2+ free treatment was decreased, either in neurons cultured for 6 d or neurons cultured for 17 d, and this effect showed no difference between neurons cultured for 6 d and 17 d; Pre and co treated with 10 mg/L IL 1ra NR2A mRNA expression increase induced by Mg 2+ free treatment in neurons cultured for 17 d was decreased, and NR2A mRNA expression showed no difference between IL 1ra group and age matched control group, but have no effect on neurons cultured for 6 d; Pre and co treated with 10 mg/L IL 1ra have NR2B mRNA expression increase induced by Mg 2+ free treatment was not affected, either in neurons cultured for 6 d or neurons cultured for 17 d. Conclusion:Neuroprotection of IL 1Ra in seizure induced injury is age dependent. The mech anism of the neuroprotection of IL
出处
《北京大学学报（医学版）》
CAS
CSCD
北大核心
2004年第4期374-378,共5页
Journal of Peking University:Health Sciences
基金
国家自然科学基金资助项目 ( 3 990 0 160
3 0 2 70 44 8)~~
