目的研究氧化槐定碱(OSR)对原代培养新生大鼠海马神经元氧糖剥夺再灌注损伤后Glu含量与NR1表达的影响。方法以原代培养的新生大鼠海马神经元为研究对象,建立氧糖剥夺再灌注损伤模型。采用化学比色法测定神经细胞培养液中谷氨酸(Glu)的含量,Western blot方法和实时荧光定量PCR技术检测大鼠海马神经细胞NMDA受体NR1亚基蛋白和mRNA的表达。结果与氧糖剥夺再灌注损伤组比较,OSR治疗组(20、5、1.25 mg·L-1)可减少神经细胞培养液中Glu的含量(P〈0.05);OSR治疗组(20μg·L-1)可明显抑制NMDA受体NR1亚基蛋白和mRNA的表达(P〈0.05)。结论 OSR通过降低Glu含量和减少NR1表达,减轻兴奋性氨基酸毒性,对新生大鼠海马神经元缺氧损伤具有明显的保护作用。
Objective To investigate the effects of OSR on content of glutamate and expressions of NR1 in neonatal rat primary cultured hippocampal neurons injured by oxygen- glucose deprivation and reperfusion. Methods Anoxia injury of hippocampal neurons that cultured in vitro were produced by physical oxygen deficiency and using culture fluid without Glu. The culture medium glutamate content was measured by chemical colorimetry. Western blotting assay and real time quantitative PCR assay were used to evaluate the expressions of NMDARNR1 protein and mRNA. Results Compared with the OGD / Rep group,treatment with OSR( 20,5,1. 25 mg· L- 1) significantly inhibited Glutamate elevation( P 0. 05,0. 01). Compared with the OGD/Rep group,treatment with OSR( 20 mg·L- 1) decreased the expressions of NMDANR1 protein and mRNA( P 0. 05,0. 01). Conclusion OSR could reduce pathologic injury by decreasing or reducing contents of glutamate and expressions of NMDARNR1,and has significantly protective effects on OGD / Rep- induced neuronal damages in rat primary neuron cultures.