远程缺血后处理抑制心肺复苏后大鼠海马神经细胞自噬  

Effect of remote ischemic postconditioning on autophagy of hippocampal neural cells after cardiopulmonary resuscitation in rats

作　　者：谢彪 高旭辉 黄洋 周翔 谭智天 朱水波 XIE Biao;GAO Xu-hui;HUANG Yang;ZHOU Xiang;TAN Zhi-tian;ZHU Shui-bo(Department of Thoracic Cardiovascular Surgery, General Hospital of Central Theater Command , Wuhan 430070 , China;partment of Anesthesiology, General Hospital of Central Theater Command , Wuhan 430070 , China)

机构地区：[1]中国人民解放军中部战区总医院心胸外科,湖北武汉430070 [2]中国人民解放军中部战区总医院麻醉科,湖北武汉430070

出　　处：《中国病理生理杂志》2019年第9期1625-1629,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81471831)

摘　　要：目的:观察远程缺血后处理(RIPostC)对心肺复苏后(CPR)大鼠海马神经细胞自噬水平的影响。方法: 45只雄性SD大鼠随机分为假手术组(sham组)、心脏停搏(CA)/CPR组和RIPostC组,每组15只。通过夹闭气管导管建立窒息性CA/CPR模型。在自主循环恢复(ROSC)后不同时点进行神经功能缺损评分(NDS),ROSC后24 h处死大鼠并取出海马组织,Western blot法检测海马神经细胞LC3-Ⅱ/LC3-Ⅰ和beclin-1蛋白的表达,TUNEL法检测细胞凋亡,免疫荧光观察LC3颗粒形成,透射电镜观察细胞自噬小体和线粒体的超微结构形态学变化。结果:与sham组相比,CA/CPR组NDS评分降低,LC3-Ⅱ/LC3-Ⅰ和beclin-1蛋白表达均升高( P <0.05),神经细胞凋亡增多( P <0.05);与CA/CPR组相比,RIPostC组的NDS评分升高,LC3-Ⅱ/LC3-Ⅰ和beclin-1表达下降( P <0.05),神经细胞凋亡减少( P <0.05),LC3颗粒形成减少,细胞内自噬小体减少,线粒体结构损伤减轻。结论:远程缺血后处理可改善心肺复苏后大鼠神经功能,这可能与抑制海马神经细胞过度自噬有关。AIM: To observe the effect of remote ischemic post-conditioning (RIPostC) on autophagy of hippocampal neural cells after cardiopulmonary resuscitation (CPR) in rats. METHODS: Male SD rats ( n =45) were randomly divided into sham operation group (sham group), cardiac arrest (CA)/CPR group and RIPostC group, with 15 rats in each group. A CPR model of asphyxiated CA was established by clamping the tracheal tube. Neurological deficit scoring (NDS) was performed at different time points after return of spontaneous circulation (ROSC). The rats were sacrificed 24 h after ROSC and hippocampal tissues were removed. Western blot was used to detect autophagy markers LC3-II/LC3-I and beclin-1 in the hippocampal tissues. The apoptosis was detected by TUNEL method. The formation of LC3 particles was observed by immunofluorescence. The ultrastructural changes of autophagosomes and mitochondria were observed under transmission electron microscope. RESULTS: Compared with sham group, the NDS scores of CA/CPR group were decreased, the protein expression of LC3-II/LC3-I and beclin-1 was increased ( P <0.05), and the apoptosis of the neural cells was increased ( P <0.05). Compared with CA/CPR group, the NDS scores in RIPostC group was increased, the protein expression of LC3-II/LC3-I and beclin-1 was decreased ( P <0.05), and the neural cell apoptosis was decreased ( P <0.05). The number of LC3 particles was decreased, intracellular autophagosome number was reduced, and the mitochondrial structure damage was alleviated. CONCLUSION: Remote ischemic post-conditioning improves neurological function in rats after CPR, which may be related to inhibition of excessive autophagy in hippocampus.

关 键 词：心脏停搏 心肺复苏 海马 自噬 远程缺血后处理 

分 类 号：R541.7[医药卫生—心血管疾病] R363[医药卫生—内科学]