K+ATP通道开放剂尼可地尔对缺血性脑卒中小鼠模型的脑保护作用及机制  

The neuroprotective effects and mechanisms of nicorandil,a K+ATP-channel opener,on mice with ischemic stroke

作　　者：付孝娟 赵源征[1] FU Xiaojuan;ZHAO Yuanzheng(Department of Neurology,the Fifth Affiliated Hospital of Zhengzhou University,Zhengzhou Henan 450052,China)

机构地区：[1]郑州大学第五附属医院神经内科,450052

出　　处：《中国神经免疫学和神经病学杂志》2023年第1期25-29,38,共6页Chinese Journal of Neuroimmunology and Neurology

基　　金：河南省高等学校重点科研项目计划(编号:22B320017);2022年精尖、冷门、绝学学科支持计划(编号:XKLMJX202211)。

摘　　要：目的探讨K+ATP开放剂尼可地尔对缺血性脑卒中小鼠的脑保护作用及机制。方法选择雄性KM小鼠72只,将小鼠随机分为假手术组、大脑中动脉闭塞(middle cerebral artery occlusion,MCAO)模型组、尼可地尔干预组,每组各24只。采用改良线栓法建立MCAO小鼠模型。尼可地尔干预组小鼠于造模术后按体重7.5 mg/(kg·d)给予尼可地尔灌胃干预,余小鼠给予等量生理盐水灌胃。造模后3 d处死小鼠,测量各组小鼠的脑含水量;采用TTC染色法测定脑梗死体积;采用免疫荧光法观察缺血半暗带区Iba-1^(+)/NLRP3^(+)共表达细胞数(Iba-1标记小胶质细胞,NLRP3标记焦亡炎症小体);采用Western Blot分析脑梗死组织核苷酸结合寡聚化结构域样受体蛋白3(NOD-like receptor protein 3,NLRP3)、活化的半胱氨酸天冬氨酸蛋白水解酶-1(cleaved cysteinyl aspartate specific proteinase-1,cleaved-caspase-1)、白细胞介素18(interleukin-18,IL-18)和IL-1β蛋白的相对表达水平。结果与MCAO模型组比较,尼可地尔干预组小鼠脑含水量明显降低,脑梗死体积减小,梗死缺血半暗带区Iba-1^(+)/NLRP3^(+)共表达细胞数量降低,脑梗死区NLRP3、cleaved-caspase-1、IL-18和IL-1β焦亡相关蛋白相对表达水平降低(均P<0.05)。结论尼可地尔对缺血性脑卒中小鼠具有脑保护作用,其机制可能与尼可地尔抑制小胶质细胞焦亡和减轻炎症反应有关。Objective To investigate the neuroprotective effect of K-ATP opener nicorandil on ischemic stroke mice and its mechanisms.Methods Seventy-two male KM mice were randomly divided into three groups equally:sham group,middle cerebral artery occlusion(MCAO)model group and nicorandil intervention group.The MCAO mouse model was established by the modified suture occlusion method.The mice in the nicorandil intervention group were given intragastric administration of nicorandil with 7.5 mg/(kg·d)after modeling,and other mice were given the same amount of normal saline by gavage.The mice were sacrificed at 3 days after modeling.We observed the changes of brain water content in each group of mice.Cerebral infarct volume was measured by TTC staining.The expression of Iba-1^(+)/NLRP3^(+)cells around cerebral infarction was detected by immunofluorescence(Iba-1 labeled microglia,NLRP3 labeled pyroptosis inflammasome).The relative expression levels of NOD-like receptor protein 3(NLRP3),cleaved cysteinyl aspartate specific proteinase-1(cleaved-caspase-1),interleukin-18(IL-18)and interleukin-1β(IL-1β)proteins in cerebral infarction tissue were analyzed by Western Blot.Results Compared with the MCAO model group,the brain water content and infarct volume of nicorandil intervention group were significantly decreased,and the number of Iba-1^(+)/NLRP3^(+)co-expressing cells in ischemic penumbra area of infarction was decreased.The relative expression levels of NLRP3,cleaved-caspase-1,IL-18 and IL-1βpyroptosis related proteins in cerebral infarction area were decreased(all P<0.05).Conclusions Nicorandil has a protective effect on ischemic stroke mice and the mechanism may be related to inhibiting microglia pyroptosis and reducing inflammatory responses.

关 键 词：尼可地尔 脑缺血 再灌注损伤 小胶质细胞 焦亡 炎症 

分 类 号：R743.33[医药卫生—神经病学与精神病学]