亚低温对脑缺血再灌注损伤的治疗作用及其机制  

Therapeutic effect of mild hypothermia on cerebral ischemia-reperfusion injury and its molecular mechanism

作　　者：韩刚 王载忠 张磊[2] Han Gang;Wang Zaizhong;Zhang Lei(Department of Neurology,Zhumadian Central Hospital,Zhumadian 463000,China;ICU,Department of Neurosurgery,Hospital of Cerebrovascular Disease,Henan People’s Hospital,Zhengzhou 450000,China)

机构地区：[1]黄淮学院附属驻马店市中心医院神经外一科,463000 [2]河南省人民医院脑血管病医院神经外科重症监护室,郑州450000

出　　处：《中华实验外科杂志》2022年第9期1662-1665,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨亚低温对脑缺血再灌注损伤的治疗作用及其分子机制。方法30只SD大鼠随机分为假手术组、模型组和亚低温组,亚低温组大鼠采用头部亚低温(33℃~35℃)预处理3 h,假手术组和模型组不做处理。采用苏木精-伊红(HE)染色分析大鼠脑组织形态变化;采用氯化三苯基四氮唑(TTC)染色法分析梗死面积;采用原位缺口末端标记法(TUNEL)染色法测定大鼠脑组织细胞细胞凋亡;采用荧光定量聚合酶链反应(PCR)分析自噬相关基因Beclin1、Atg7和Atg5表达水平;采用蛋白质印迹法(Western blot)分析自噬底物p63和LC3-Ⅱ表达水平。计量数据采用单因素方差分析。结果亚低温组大鼠神经功能评分[(1.90±0.74)分]明显低于模型组[(3.40±0.70)分],差异有统计学意义(t=4.666,P<0.05)。亚低温组大鼠运动功能评分[(6.40±0.97)分]明显低于模型组[(3.60±1.17)分],差异有统计学意义(t=5.846,P<0.05)。亚低温组大鼠脑组织梗死百分比[(24.79±2.57)%]明显低于模型组[(46.91±5.79)%],差异有统计学意义(t=4.855,P<0.05)。亚低温组大鼠脑组织细胞凋亡比例[(28.04±7.19)%]明显低于模型组[(63.76±5.79)%],差异有统计学意义(t=12.110,P<0.05)。亚低温组大鼠脑组织Beclin1和Atg7 mRNA表达水平(1.96±0.08;1.83±0.14)明显高于模型组(1.44±0.08;1.65±0.97),差异有统计学意义(t=14.550、9.849,P<0.05)。亚低温组大鼠脑组织p62和LC3-Ⅱ相对表达水平(1.40±0.09、1.39±0.11)明显低于模型组(0.84±0.04、0.88±0.07),差异有统计学意义(t=18.200、12.210,P<0.05)。结论亚低温处理可显著促进脑组织细胞自噬基因表达,促进细胞自噬,对脑缺血再灌注损伤起到保护作用。Objective To investigate the therapeutic effect of mild hypothermia on cerebral ischemia-reperfusion injury and its molecular mechanism.Methods A total of 30 SD rats were randomly divided into sham operation group,model group and mild hypothermia group.The rats in mild hypothermia group were pretreated with mild hypothermia(33℃-35℃)at head for 3 h.The rats in sham operation group and model group were not treated.Hematoxylin and eosin(HE)staining was used to analyze the morphological changes of rat brain.The infarct size was analyzed by tovanto transit commission(TTC)staining.The apoptosis of rat brain cells was tested by terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)staining.The expression levels of autophagy related genes Beclin1,Atg7 and ATG5 were analyzed by fluorescence quantitative polymerase chain reaction(PCR).The expression levels of autophagy substrates p62 and LC3-Ⅱwere analyzed by Western blotting.Results The neurological function score in mild hypothermia group[(1.90±0.74)points]was significantly lower than that in model group[(3.40±0.70)points,t=4.666,P<0.05].The motor function score in mild hypothermia group[(6.40±0.97)points]was significantly lower than that in model group[(3.60±1.17)points,t=5.846,P<0.05].The percentage of cerebral infarction in mild hypothermia group[(24.79±2.57)%]was significantly lower than that in model group[(46.91±5.79)%,t=4.855,P<0.05].The apoptosis rate in mild hypothermia group[(28.04±7.19)%]was significantly lower than that in model group[(63.76±5.79)%](t=12.110,P<0.05).The expression levels of Beclin1 and Atg7 mRNA in the brain tissue in mild hypothermia group(1.96±0.08,1.83±0.14)were significantly higher than those in the model group(1.44±0.08,1.65±0.97,t=14.550,9.849,P<0.05).The relative expression levels of p62 and LC3-Ⅱin the brain tissue in mild hypothermia group(1.40±0.09,1.39±0.11)were significantly lower than those in the model group(0.84±0.04,0.88±0.07,t=18.200,12.210,P<0.05).Conclusion Mild hypothermia treatment

关 键 词：亚低温 脑缺血再灌注损伤 凋亡 自噬 

分 类 号：R743[医药卫生—神经病学与精神病学]