NK细胞活化性受体及其配体在缺血性脑卒中模型小鼠脑组织中的表达变化  

Expression Change of NKG2D Receptor and Its Ligands in Ischemic Brain Tissue of Model Mice

作　　者：陈秋竹 杜欧 朱丽 曹丽丽[2] 杜俊蓉[1] Chen Qiuzhu;Du Ou;Zhu Li;Cao Lili;Du Junrong(West China School of Pharmacy, Sichuan University, Chengdu 610041, China;School of Preclinical Medicine, Chengdu University, Chengdu 610106, China)

机构地区：[1]四川大学华西药学院,成都610041 [2]成都大学基础医学院,成都610106

出　　处：《成都医学院学报》2022年第3期288-292,共5页Journal of Chengdu Medical College

基　　金：四川省科技厅科技计划重点研发项目(No:2018SZ0205)。

摘　　要：目的探讨NK细胞活化性受体(NKG2D)信号通路在缺血脑组织中的表达及其在缺血性脑损伤中的潜在作用。方法采用双侧颈总动脉持久性结扎法(2VO),构建C57BL/6雄性小鼠全脑缺血再灌注损伤模型组(2VO组)和假手术对照组(Sham组),每组12只。脑缺血20 min、再灌注72 h后,处死小鼠收集大脑,Nissl染色观察海马CA1区和纹状体CPu区神经元的损伤程度,实时荧光定量PCR法和蛋白质印迹技术检测缺血脑组织中NKG2D及其配体(H60、Rae-1、Mult1)的表达,实时荧光定量PCR法检测NKG2D受体下游接头蛋白DAP10及促炎因子(TNF-α、IL-6、IL-1β、iNOS)的mRNA表达。结果Nissl染色结果显示,2VO组小鼠海马CA1区和纹状体CPu区的神经元严重损伤(P<0.001);2VO组小鼠缺血脑组织中NKG2D及其3种配体的mRNA水平和蛋白表达水平均高于Sham组(P<0.05);下游接头蛋白DAP10以及促炎因子的mRNA水平均明显高于Sham组(P<0.05)。结论脑缺血介导的NKG2D受体信号通路激活可能在小鼠脑缺血损伤炎症反应中发挥重要作用。Objective To explore the expression of NK cell activating receptor natural killer group 2 member D(NKG2D)signaling pathway in ischemic brain tissue and its potential role in ischemic brain injury.Methods The common carotid artery was ligated bilaterally to construct a global cerebral ischemia-reperfusion injury model(2VO)in C57BL/6 male mice.A sham operation control group(Sham)was also constructed.There were 12 mice in each group.After 20 minutes of cerebral ischemia and 72 hours of reperfusion,mice were sacrificed and the brains were collected.Nissl staining was used to observe the damage degree of neurons in hippocampal CA1 and striatal CPu subregions,respectively.The qRT-PCR and Western Blot were utilized to detect the expression of NKG2D and its ligands(H60,Rae-1,Mult1).The qRT-PCR was utilized to detect the mRNA expression of NKG2D receptor adaptor protein DAP10,and pro-inflammatory factors(TNF-a,IL-6,IL-1β,iNOS)in ischemic brain tissue.Results Nissl staining showed that the neurons in the hippocampal CA1 area and striatal CPu area of 2VO mice were severely damaged(P<0.001).The mRNA and protein levels of NKG2D and its three ligands in the ischemic brain tissue of the 2VO mice were significantly higher than those of the Sham group(P<0.05).The mRNA levels of adaptor protein DAP10,as well as pro-inflammatory factors in the ischemic brain tissue of the 2VO mice were significantly higher than those of the Sham group(P<0.05).Conclusion Cerebral ischemia-mediated activation of NKG2D receptor signaling pathway may play an important role in the inflammatory response of ischemic brain injury in mice.

关 键 词：脑缺血 NK细胞活化性受体 NK细胞活化性受体配体 神经炎症 

分 类 号：R741.02[医药卫生—神经病学与精神病学]