亚低温对心肺复苏后大鼠海马CA1区神经元细胞自噬的影响  

Effects of mild hypothermia on autophagy in hippocampal CA1 neurons in rats after CPR

作　　者：孙占朋 廖晓星[1] 尹美娴 杨焰[1] 魏红艳[1] 黎博[1] 荆小莉[1] 胡春林[1] 

机构地区：[1]中山大学附属第一医院急诊科,广州510080 [2]广东药科大学生命科学与生物制药学院,广州510006

出　　处：《中华急诊医学杂志》2017年第5期549-553,共5页Chinese Journal of Emergency Medicine

基　　金：国家自然科学基金（81372021）

摘　　要：目的观察亚低温对自主循环恢复后（ROSC）大鼠海马CA1区神经元细胞自噬的影响。方法36只Wistar大鼠随机（随机数字法）分为常温治疗组和亚低温治疗组，心外膜致颤法建立大鼠心脏骤停（cardiac arrest, CA）心肺复苏模型，ROSC后2 h和4 h每组各处死3只动物取大脑皮层，Western blot方法观察p-AMPK和自噬标志物Beclin-1、LC3的表达，免疫荧光观察LC3颗粒的形成。ROSC后24、48和72 h分别进行神经功能缺损（neurologic deficit score，NDS）评分，ROSC后72 h处死动物取大脑进行TUNEL染色计算海马CA1区神经元细胞凋亡指数。结果常温组ROSC后2 h和4 h p-AMPK、Beclin-1表达水平以及LC3-Ⅱ/LC3-Ⅰ比例均低于Hypothermia组（P〈0.05）;亚低温组CA1区神经元胞浆内可见明显的LC3颗粒形成；ROSC后24 h、48 h、72 h常温组和亚低温组大鼠的NDS评分分别为320 vs. 205、285 vs. 140、266 vs. 120，ROSC后72 h常温组CA1区神经元细胞凋亡指数高于亚低温组（P〈0.05）。结论心肺复苏后亚低温治疗可促进大鼠海马神经元细胞自噬，减少神经元细胞凋亡。Objective To establish the cardiac arrest-cardiopulmonary resuscitation model in rats, and to observe the effect of mild hypothermia on autophagy in hippocampal CAI neurons after ROSC. Methods A total of 36 Wistar rats were randomly divided into two groups ： normal temperature treatment group （NT group） and mild hypothermia treatment group （ HT group）. To establish the cardiac arrest- cardiopulmonary resuscitation （CA-CPR） model in rats by epicardial electrical stimulation induced ventricular fibrillation, and to sacrifice 3 animals in each group to obtain the brain cortex in 2nd and 4th hours after ROSC in order to observe the expression of p-AMPK by electron microscope and LC3 granules through Western blot. The neurological deficit score （NDS） was assessed in 24, 48, 72 hours respectively after ROSC. To sacrifice the animals so as to take the cerebrum in 72 hours after ROSC, then calculate the apoptotic index of the hippocampal CA1 neurons, which were dyed through TUNEL method. Results The expression of p-AMPK, Beclin-1 and LC3- Ⅱ/LC3- Ⅰ ratio in Normothermia group were all lower than the Mild hypothermia group （ P 〈 0. 05 ）, the neurons plasma of hippocampal CA1 area in the Hypothermia group demonstrated obvious LC3 granules formation, the NDS score of the Normothermia group and the Mild hypothermia group in ROSC24h, ROSCA8h, ROSC72h were 320vs205, 285vs140, 266vs120,respectively. The apoptotic index of the hippocampal CA1 area in the Normothemia group in ROSC72h was higher than the Mild hypothermia group, （P 〈 0. 05 ）. Conclusions Mild hypothermia after cardiopulmonary resuscitation womotes autophagy of the hippocampal CA1 area neurons in rats and reduce neuronal apoptosis.

关 键 词：心搏骤停 心肺复苏 亚低温 大鼠 自噬 

分 类 号：R459.7[医药卫生—急诊医学]