低温联合褪黑素降低小鼠局灶性脑缺血再灌注损伤  

Hypothermia Combined with Melatonin Decreases Reperfusion Injuries of Mouse Focal Cerebral Ischemia

作　　者：陈武[1] 李毅[1] 段发兰[1] 李国玲[1] 陈琴华[2] 张正洪[3] 

机构地区：[1]湖北医药学院附属东风医院检验部,湖北十堰442008 [2]湖北医药学院附属东风医院实验中心,湖北十堰442008 [3]湖北医药学院附属东风医院神经内科,湖北十堰442008

出　　处：《湖北医药学院学报》2017年第1期1-5,F0002,共6页Journal of Hubei University of Medicine

基　　金：国家自然科学基金项目(81344438)资助

摘　　要：目的:调查物理性低温(physical hypothemia,PH)联合褪黑素(melotonin,MLT)对小鼠脑缺血再灌注损伤的保护作用。方法:以H2O2作用C57BL/6小鼠原代皮层神经元(primary cortical neuron,PCN)制备体外氧化应激模型;同时以线栓阻断法制作小鼠中脑动脉阻塞(middle cerebral artery occlusion,MCAO)再灌注模型;细胞动物实验均设PH(34℃)组、MLT组、PH(34℃)+MLT组。对PCN的保护作用采用4',6-二脒基-2-苯基吲哚(4',6-diamidino-2-phenylindole,DAPI)染色、乳酸脱氢酶(lactate dehydrogenase,LDH)活性评价;MCAO再灌注动物采用2,3,5-氯化三苯基四氮唑(2,3,5-triphenyltetrazolium chloride,TTC)染色、神经行为学评价。结果:H2O2、PH(34℃)、MLT、PH(34℃)+MLT作用于PCN的相对LDH活性分别为:172.8%±12.1%、150.3%±4.4%、141.6%±9.3%、117.2%±8.2%,各组间有显著差异(F=33.10,P=0.000);与对照组相比,PH(34℃)组、MLT组脑相对梗死体积分别减少了14.7%(t=4.02,P=0.008)、26.8%(t=7.89,P=0.000),而联合处理组减少了56.9%(t=18.46,P=0.000);三组神经行为学评分也较对照组有明显改善。结论:低温、褪黑素对小鼠局灶性脑缺血再灌注损伤均有保护作用,而联合使用效果更明显。Objective To explore the protective effects of physical hypothermia（PH） combined with melatonin（MLT） on cerebral ischemia-reperfusion injuries. Methods An in vitro oxidative stress model of mouse primary cortical neuron（PCN） was prepared using hydrogen peroxide（H2O2）,and a mouse model of middle cerebral artery occlusion-reperfusion was prepared using suture embolization method. Both cellar and animal model studies included PH（34 ℃） group,melatonin group,PH（34 ℃） plus melatonin group. The protective effect on PCN was evaluated by 4＇,6-diamidino-2-phenylindole（DAPI） staining and lactate dehydrogenase（LDH） activity assay,and MCAO-reperfused animals were evaluated by 2,3,5-triphenyltetrazolium chloride（TTC） staining and neurological behavior scoring. Results The relative LDH activities of PCN in H2O2,PH,MLT,PH plus MLT groups were 172.8%±12.1%,150.3%±4.36%,141.6%±9.3% and 117.2% ±8. 2% respectively,which had significant difference from each other（F = 33.10,P = 0.000）. In comparison with vehicle group,the relative brain infarct volumes of HT,MLT and combination groups were decreased by 14.7%,26.8%,56.9%,respectively. The neurobehavioral scoring of three treatment groups were also better than vehicle group. Conclusion The present results indicated that both HT and MLT pretreatment had protective effects on ischemia-reperfusion brain injuries. While combining HT with MLT could provide better protection.

关 键 词：低温 褪黑素 缺血再灌注 小鼠 中脑动脉阻塞 

分 类 号：R743.3[医药卫生—神经病学与精神病学]