摘要
目的探讨线粒体膜Kir6.1/K-ATP通道对脑缺血预处理大鼠神经功能的修复作用及作用机制。方法将72只雄性SD大鼠随机分为假手术组、模型组和缺血预处理组,每组各24只。采用Zea Longa线栓法建立大鼠脑缺血预处理模型。于造模后12、24、36和48 h时采用Zea Longa评分评价大鼠神经功能,采用MTT法检测脑梗死体积,采用Western blot法检测大鼠脑组织凋亡蛋白(包括Bax、Caspase-3、Cyt-c、Bcl-2)和Kir6.1蛋白表达,分别采用线粒体通透性转化孔检测试剂盒和线粒体膜电位试剂盒检测大鼠脑组织线粒体膜电势及线粒体膜通透性,采用荧光定量PCR测定脑组织miR-7表达水平。结果假手术组大鼠无神经功能缺损症状,造模24、36、48 h时模型组Zea Longa评分和脑梗死体积均高于缺血预处理组(P<0.05)。造模24、36和48 h时,模型组Bax、Caspase-3及Cyt-c蛋白表达水平均高于假手术组(均P<0.05),而缺血预处理组较模型组低(均P<0.05)。三组不同时间点Bcl-2蛋白表达水平比较差异无统计学意义(P>0.05)。造模24、36和48 h时,模型组Kir6.1蛋白水平、线粒体膜电位光密度及膜通透性转换孔平均光密度均低于假手术组(均P<0.05),而缺血预处理组其水平均高于模型组(均P<0.05)。造模12、24、36和48h时,模型组miR-7表达水平均低于假手术组(均P<0.05),造模48 h时,缺血预处理组miR-7表达量高于模型组(P<0.05)。结论脑缺血预处理可能通过抑制Kir6.1蛋白和miR-7过表达改善脑缺血所导致的神经功能损伤和神经元凋亡。
Objective To explore the effect and mechanism of kir6.1/k-ATP channel of mitochondrial membrane on neural function repair in rats with cerebral ischemic pretreatment.Methods 72 male SD rats were randomly divided into a sham operation group,a model group and an ischemic pretreatment group;there were 24 rats in each group.Zea longa thread embolism was used to establish the rat model of cerebral ischemic pretreatment.At 12,24,36 and 48 hours after modeling,the neural function of rats was evaluated by Zea longa score.The volume of cerebral infarction was measured by MTT method.Western blot was used to detect the expression of apoptotic proteins(including Bax,Caspase-3,Cyt-C,and Bcl-2)and Kir6.1 in rat brains.Mitochondrial permeability and membrane potential of rat brain were measured by mitochondrial permeability transition pore test kit and mitochondrial membrane potential test kit respectively.The expression level of miR-7 in brain tissue was determined by fluorescence quantitative PCR.Results There was no neurological deficit in the sham operation group.At 24,36 and 48 hours after the establishment of the model,the Zea longa score and the volume of cerebral infarction in the model group were higher than those in the ischemic pretreatment group(P<0.05).The expressions of Bax,caspase-3 and Cyt-C protein in the model group were higher than those in the sham operation group(P<0.05),while the ischemic pretreatment group was lower than the model group(P<0.05).There was no significant difference in the expression level of Bcl-2 between the three groups at different time points(all P>0.05).At 24,36 and 48 hours,Kir6.1 protein level,mitochondrial membrane potential optical density and average optical density of membrane permeability transition pore in the model group were lower than those in the sham operation group(all P<0.05),while those in the ischemic pretreatment group were higher than those in the model group(all P<0.05).The expression level of miR-7 in the model group was lower than that in the sham operation group(
作者
宋笑雨
刘开扬
薛茜
杨金水
张宇
胡雪霄
邹玉安
SONG Xiaoyu;LIU Kaiyang;XUE Qian;YANG Jinshui;ZHANG Yu;HU Xuexiao;ZOU Yu′an(不详;Department of Neurology,the First Affiliated Hospital of Hebei North University,Zhangjiakou Hebei 075061,China)
出处
《中国神经免疫学和神经病学杂志》
CAS
北大核心
2021年第1期13-19,共7页
Chinese Journal of Neuroimmunology and Neurology
基金
河北省2015年度医学科学研究重点课题项目(20150054)。
关键词
缺血预处理
脑缺血
ATP敏感性钾通道
线粒体
细胞凋亡
钾通道
内向整流
ischemic preconditioning
brain ischemia
ATP-sensitive potassium channel
mitochondria
apoptosis
potassium channels,inwardly rectifying
