氧糖剥夺对小鼠小胶质细胞NKG2D受体及其配体表达的影响  

Expression of NKG2D receptor and its ligands in mouse microglia after oxygen glucose deprivation injury

作　　者：刘琳 李晶莹 曹丽丽[2] 杜俊蓉[1] LIU Lin;LI Jingying;CAO Lili;DU Junrong(West China School of Pharmacy, Sichuan University, Chengdu 610041, China;Basic Medical School, Chengdu University, Chengdu 610106, China)

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

出　　处：《西部医学》2021年第11期1570-1573,共4页Medical Journal of West China

基　　金：四川省科技计划项目(2-205)。

摘　　要：目的探究自然杀伤细胞活化受体(NKG2D)在氧糖剥夺损伤小胶质细胞中的表达变化及炎症反应中的潜在作用。方法建立小鼠神经元细胞HT22氧糖剥夺(OGD)模型,造模19小时后复氧复糖,分别收集正常条件培养基(Control)和OGD培养基(Model)刺激小鼠BV2小胶质细胞。CCK-8法检测HT22和BV2细胞活性;qPCR检测BV2细胞中NKG2D受体信号通路及炎症因子表达;ELISA法检测BV2细胞培养上清中炎症因子含量。结果OGD可介导HT22神经元细胞存活率明显下降(P<0.01),但OGD上清对BV2细胞活性无明显影响(P>0.05)。与Control组比较,NKG2D受体及其H60配体、接头蛋白DAP10的mRNA表达水平明显上升(P<0.01),但RAE-1与MULT1配体未见明显变化(P>0.05);同时,肿瘤坏死因子(TNF-α)在Model组BV2细胞中的mRNA水平及培养基中的蛋白含量均明显上升(P<0.01)。结论氧糖剥夺可诱导BV2细胞H60/NKG2D、下游效应器DAP10及炎症因子表达。提示NKG2D受体信号通路可能在脑缺血后小胶质细胞的炎症反应中发挥重要作用。Objective To investigate the expression changes of Natural Killer Group 2 member D(NKG2D)signaling pathway in microglia of mice after oxygen-glucose deprivation injury and its potential role in microglia inflammatory response.Methods A mouse neuronal HT22 oxygen-glucose deprivation(OGD)Model was established(HT22-model group).After 19 hours of modeling,the cells were oxygenated and glycosylated,and normal conditioned media(Control)and OGD media(Model)were collected to stimulate mouse BV2 microglia cells.The cell viability of HT22 and BV2 cells was detected by CCK-8 assay.NKG2D receptor signaling pathway and inflammation mediators expression in BV2 cells were detected by qPCR.The content of inflammation mediators in supernatant of BV2 cell culture was determined by ELISA.Results The survival rate of HT22 neurons was significantly decreased by OGD(P<0.01),but the cell viability of BV2 cells was not affected by OGD supernatant(P>0.05).Compared with the Control group,the mRNA expression levels of NKG2D receptor,its H60 ligand and adaptor protein DAP10 were significantly increased(P<0.01),but there were no significant changes in RAE-1 and MULT1 ligand(P>0.05).Meanwhile,the mRNA level and the protein content of BV2 cells of tumor necrosis factor(TNF-α)in Model group were significantly increased(P<0.01).Conclusion Oxygen-glucose deprivation can induce the expression of H60/NKG2D,downstream effector DAP10 and inflammation mediators in BV2 cells.These results suggest that the NKG2D receptor signaling pathway may play an important role in the inflammatory response of microglia after cerebral ischemia.

关 键 词：氧糖剥夺 小胶质细胞 NKG2D受体及其配体 神经炎症 缺血性脑卒中 

分 类 号：R743.31[医药卫生—神经病学与精神病学]