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心肺复苏大鼠神经元凋亡和内质网应激相关因子 GRP78及CHOP表达的变化
赵芯晨王晶何婧瑜周小超田欣
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摘要:
目的 研究心跳骤停复苏后大鼠脑内质网应激相关因子葡萄糖调节蛋白78(GRP78)和 CCAAT/增强子结合蛋白同源蛋白(CHOP)表达的动态变化及其与神经元凋亡的关系,从而探讨内质网 应激与脑缺血再灌注损伤的机制。方法 将48只SD大鼠随机分为对照组和复苏组,采用经食道起搏诱 发室颤法制备大鼠心肺复苏模型;应用尼氏染色观察各组大鼠海马CA1区6 h、12 h、24 h、48 h存活神经 元形态;应用TUNEL染色法,观察各组大鼠海马CA1区神经元凋亡情况,并进行凋亡阳性细胞计数;应 用免疫组化法检测GRP78及CHOP抗原的表达、应用Western Blot法检测GRP78及CHOP蛋白的表达量。 结果 与对照组比较,复苏组大鼠海马CA1区神经元细胞变性更严重,出现凋亡细胞,随着再灌注时间 的延长,凋亡细胞明显增多。与对照组比较,复苏组GRP78阳性细胞于再灌注 6 h逐渐增多,于12 h明显 增高,此后表达逐渐减少;复苏组CHOP于再灌注6 h开始阳性细胞即有表达,逐渐增多,于再灌注 24 h 显著增加,直至48 h。结论 心跳骤停复苏后诱发内质网应激,早期通过上调促生存因子GRP78蛋白的 表达达到自我保护作用,晚期通过上调凋亡因子CHOP诱导神经元凋亡。
关键词:  心肺复苏  内质网  神经元  凋亡  GRP78  CHOP
DOI:10.3969/j.issn.1009-6574.2017.10.004
基金项目:北京市医管局重点医学专业发展项目(ZYLX201706)
Changes of neuronal apoptosis and expression of endoplasmic reticulum stress related factors GRP78and CHOP in rats after cardiopulmonary resuscitation
()
Abstract:
Objective To investigate the dynamic changes of endoplasmic reticulum stress-related factors, the expression of glucose regulated protein 78( GRP78) and CCAAT/ enhancer binding protein homologue protein( CHOP) in rats after cardiac arrest and resuscitation, their relationship with neuronal apoptosis, so as to explore the mechanism of endoplasmic reticulum stress and cerebral ischemia-reperfusion injury. Methods A total of 48 male SD rats were randomly divided into two groups: the group of without cardiopulmonary resuscitation( CPR)( control group)( n=24) and the group of normothermia after CPR( CPR group)( n=24). The CPR model in rats were prepared by esophagus pacing induced ventricular fibrillation. At the time of 6 h, 12 h, 24 h, and 48 h after restoration of spontaneous circulation, the morphology of the surviving neurons in the hippocampal CA1 region of rats was observed by Nissl staining. TUNEL staining was used to observe the apoptosis of hippocampal CA1 neurons in rats and to count the number of positive apoptotic cells. The expression of antigen of GRP78 and CHOP was detected by immunohistochemical method. The expression of GRP78 and CHOP was detected by Western Blot method. Results Compared with control group, the neuronal cell degeneration in the hippocampal CA1 area was more severe in the resuscitation group. Apoptotic cells increased significantly with the increase of reperfusion time. Compared with control group, GRP78 positive cells in the resuscitation group increased gradually at reperfusion 6 h, and increased significantly in 12 h, and then the expression decreased gradually. In the CPR group, CHOP positive cells began to be expressed at the beginning of the reperfusion 6 h, and gradually increased, and increased significantly at 24 h until 48 h. Conclusions Endoplasmic reticulum stress is induced after CA/CPR. It can protect itself at the early stage by upregulated the expression of GRP78 protein, and apoptosis is induced by upregulated apoptosis factor CHOP at the late stage
Key words:  Cardiopulmonary resuscitation  Endoplasmic reticulum  Neurons  Apoptosis  GRP78  CHOP

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