新生大鼠缺氧缺血再灌注后海马神经元凋亡与c-Jun蛋白的表达检测

Detection of c-Jun and apoptosis in hippocampus of neonatal rats after hypoxic-ischemia brain damage and reperfusion

目的检测新生大鼠缺氧缺血再灌注后即刻早期基因c-jun的表达与海马神经元的凋亡。方法49只7d龄SD大鼠经弹性管穿线阻断右颈总动脉3h,予低氧1h,制备缺氧缺血脑损伤(HIBD)模型,并于再灌注3h、6h、12h、24h、48h、96h、7d处死大鼠,取脑组织。采用免疫组织化学SABC法检测海马CA1、CA3区c-Jun蛋白的表达。采用TUNEL染色法计数海马CA1、CA3区凋亡神经元。8只7d龄SD大鼠仅手术不行HIBD及再灌注(对照组)。结果海马CA1、CA3区c-Jun的表达和凋亡细胞数于HIBD再灌注3h、6h、12h、24h、48h、96h均高于对照组(P<0.05),c-Jun的表达于再灌注6h达高峰,锥体神经元凋亡于再灌注24h达高峰。再灌注7d组c-Jun蛋白的表达及凋亡细胞数与对照组相比差异均无统计学意义。结论c-Jun可能参与轻度HIBD后神经元的凋亡与修复。

Aim ： To detect the expression of c-Jun and cell apoptosis in hippocampus of neonatal rats after hypoxic-ischemia brain damage（HIBD） and reperfusion. Methods： A total of 49 seven day old SD rats were established HIBD model and allocated into reperfusion 3 h, 6 h, 12 h, 24 h, 48 h, 96 h, and 7 d groups. Eight rats were only operated but not established HIBD model or reperfusion（ control group）. At different time points, the rats were killed and the specimens from fight hippocampus were prepared. The protein of c-Jun was detected using immunohistochemistry. The apoptotic cells were counted using TUNEL staining. Results： In reperfusion 3 h, 6 h, 12 h, 24 h, 48 h, and 96 h groups, the expression of c-Jun and the number of apoptotic cells were higher than those in control group （ P 〈 0.05 ）. The expression of c-Jun reached peak after 3 h reperfusion, and the number of apoptotic cells reached peak after 24 h reperfusion. After 7 days reperfusion, the expression of c-Jun and the number of apoptotic cells among reperfusion groups and control group had no significant differences. Conclusion ： c-Jun may participate in the apoptosis and repairment of neurons during HIBD and reperfusion.