摘要
目的探讨2-脱氧葡萄糖(2-deoxyglucose,2-DG)对内质网应激介导的宫内窘迫胎鼠脑损伤的保护机制。方法制备胎鼠宫内窘迫模型;实验动物随机分为正常组、缺血再灌注组、假手术组及治疗组;取胎鼠脑组织行TUNEL染色,分析神经元凋亡情况;免疫组化法检测海马回CA1区GRP78及caspase-9,-12蛋白的表达;并观察2-DG对上数指标的影响。结果缺血再灌注组再灌注3hTUNEL阳性细胞数目明显增多;至再灌注24h阳性神经元数量达最大值。治疗组海马回CA1区典型TUNEL阳性神经元数量与缺血再灌注组各相应时间点比较明显减少(P<0.05)。缺血再灌注组GRP78,caspase-9,-12蛋白表达高于正常及假手术组。GRP78于再灌注3h达峰值,6h内无明显变化,以后随时间的延迟逐渐下降;caspase-12在3h内表达急剧上升,以后趋势变缓,至12h达高峰;caspase-9则于3h内无明显变化,之后迅速增加,至12h达到最大值。治疗组海马回CA1区神经元GRP78蛋白表达均显著高于缺血再灌注组,而神经元凋亡数量及caspase9,-12蛋白表达显著低于缺血再灌注组(P<0.01)。结论宫内窘迫所致的胎鼠脑组织缺血缺氧启动了内质网应激,内质网相关性死亡途经是胎鼠神经元损伤的又一机制;2-DG通过上调内质网分子伴侣GRP78的表达而对胎鼠缺血缺氧性脑损伤有明确的保护作用。
[ Objective ] To investigate the protective mechanism of 2-deoxyglucose （2-DG）in fetal rat＇s hypoxicischemic cerebral damage mediated by ER stress of intrauterine distress. [Methods] Fetal rat intrauterine distress model was set up. The pregnant rats were randomly divided into four groups： normal group, sham operated group, ischemia-reperfusion （IR） group and treatment group. The insitu end-labeled DNA （TUNEL） was used to analyze neuron apoptosis. The expression of caspase-9, -12 and GRP78 protein in hippocampus CA1 area were detected by immunohistochemistry staining. And the influence of 2-DG on these markers was observed. [Results] The number of TUNEL positive neuron in hippocampus CA1 area at 3h in the IR group increased significantly. It reached the maximum value at 24 h. On the contrary, the number of positive neuron in the 2-DG treatment group on corresponding time was lower than that in the IR group （P 〈0.05）. The expression of GRP78, caspase-9 and caspase-12 genes in IR group were, significantly higher than that of normal group and sham operated group. The expression of GRP78 increased rapidly within 3 hours after the reperfusion. It retained this level in 6 hours. Subsequently, it increased gradually. The intensity of caspase-12 was significantly increased in 3 hours. Furthermore, caspase-9 increased little within 3 hours. Subsequently, its expression increased rapidly.caspase-12 increased significantly in 12 hours. The expression of GRP78 in hippocampus CA1 area was higher than IR group on the same time. Oppositely, the apoptotic neuron and expression of caspase-9 and caspase-12 genes in 2-deoxglucose treatment group were less than that of IR group （P 〈0.01）. [Conclusions] Endoplasmic reticulum stress lead by fetal rat hypoxic-ischemic cerebral damage is induced by intrauterine distress. Endoplasmic reticulum-associated death pathway is one of the mechanisms that priming damage pathway in cerebral ischemic injury. 2-DG can protect the neuron from hypoxia-ischemia damage by promoting the expression of GRP78 in general ischemic cerebral areas, which is one of endoplasmic reticulum molecular companion.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2008年第4期432-436,共5页
China Journal of Modern Medicine
基金
重庆市自然科学基金[2004]54号80
重庆市卫生局科研基金资助项目(05-2-189)
关键词
缺血缺氧性脑损伤
内质网应激
2-脱氧葡萄糖
hypoxic-ischexnic cerebral damage
endoplasmic reticulum stress
2-deoxyglucose
