miR-331-5p靶向AQP4对大鼠星形胶质细胞中DEACMP标志物表达的影响  

Effects of miR-331-5p targeting AQP4 on the expression of DEACMP markers in rat astrocytes

作　　者：贾敏 王从平 刘慧纯 刘群会 JIA Min;WANG Cong-ping;LIU Hui-chun(Department of Neurology,Central Hospital of Enshi Tujia and Miao Autonomous Prefecture,Enshi,Hubei Province 445000,China)

机构地区：[1]恩施土家族苗族自治州中心医院神经内科,湖北恩施445000

出　　处：《解剖学研究》2021年第3期241-246,共6页Anatomy Research

基　　金：湖北省科技计划项目(2014CFB424)。

摘　　要：目的探讨一氧化碳中毒后迟发性脑病(DEACMP)标志物表达的潜在机制。方法纳入2018年7月至2020年7月于我院接受治疗的DEACMP患者36例,健康体检者32例,检测其血清中DEACMP标志物髓磷脂碱性蛋白(MBP)的表达水平。给大鼠腹膜内注射CO以建造大鼠DECAMP模型。造模成功的为DECAMP组大鼠,相应PBS处理的为对照组大鼠。水迷宫测试两组大鼠逃脱时间。苏木精伊红染色分析两组大鼠海马结构。高通量测序检测两组大鼠星形胶质细胞中的差异miRNA。过表达差异miRNA后,检测大鼠星形胶质细胞中DEACMP标志物MBP的表达水平。通过miRDB在线分析miRNA的潜在底物并通过荧光素酶报告实验验证。结果与健康体检者相比,DECAMP患者血清中MBP的表达水平升高(P<0.05)。DEACMP组中毒大鼠在第7天和第14天的逃脱时间比正常大鼠要长(P<0.05)。对照组海马的结构和细胞形状是正常的,而在DEACMP组中观察到神经元损伤,例如细胞萎缩,核固体收缩和结构模糊。与对照组相比,大鼠血清DEACMP标志物MBP的蛋白水平在第7天和第14天均上升(P<0.05)。过表达miR-331-5p时大鼠星形胶质细胞的MBP表达水平上升(P<0.05)。敲低miR-331-5p后,大鼠星形胶质细胞的MBP表达水平下降(P<0.05)。敲低AQP4时大鼠星形胶质细胞的MBP表达水平上升(P<0.05)。过表达AQP4后,大鼠星形胶质细胞的MBP表达水平下降(P<0.05)。过表达miR-331-5p后,AQP4的蛋白水平下降(P<0.05)。敲低miR-331-5p后,AQP4的蛋白水平上升(P<0.05)。miR-331-5p靶向AQP4的3端非编码区(P<0.05)。DEACMP模型中毒大鼠海马在AQP4上的阳性染色少于正常大鼠。结论miR-331-5p通过靶向AQP4mRNA的3端非编码区,减少了AQP4的蛋白表达水平,最终增加了大鼠星形胶质细胞中DEACMP标志物MBP的表达。Objective To explore the potential mechanism of delayed encephalopathy after acute carbon monoxide poisoning(DEACMP)marker expression.Methods 36 DEACMP patients and 32 healthy subjects who received treatment in our hospital from July 2018 to July 2020 were enrolled,and the serum levels of the DEACMP marker myelin basic protein(MBP)were detected.Level.CO was injected intraperitoneally to rats to build a rat DECAMP model.The rats in the DECAMP group were successfully modeled,and the rats in the control group were treated with PBS.The water maze test the escape time of the two groups of rats.Hematoxylin and eosin staining was used to analyze the hippocampal structure of the two groups of rats.High-throughput sequencing detects the differential miRNAs in astrocytes of two groups of rats.After overexpression of differential miRNA,the expression level of DEACMP marker MBP in rat astrocytes was detected.The potential substrates of miRNA were analyzed online by miRDB and verified by the luciferase report experiment.Results Compared with healthy subjects,the expression level of MBP in serum of DECAMP patients was increased(P<0.05).The escape time of poisoned rats in the DEACMP group on the 7 th and 14 th days was longer than that of the normal rats(P<0.05).The structure and cell shape of the hippocampus in the control group were normal,while neuronal damage was observed in the DEACMP group,such as cell atrophy,nuclear solid shrinkage,and structure blurring.Compared with the control group,the protein level of the serum DEACMP marker MBP increased on the 7 th day and the 14 th day(P<0.05).The MBP expression level of rat astrocytes increased when miR-331-5 p was overexpressed(P<0.05).After knocking down miR-331-5 p,the expression level of MBP in rat astrocytes decreased(P<0.05).The expression level of MBP in rat astrocytes increased when AQP4 was knocked down(P<0.05).After overexpression of AQP4,the expression level of MBP in rat astrocytes decreased(P<0.05).After overexpression of miR-331-5 p,the protein level of AQP4 decre

关 键 词：一氧化碳中毒后迟发性脑病 miR-331-5p 水通道蛋白4 星形胶质细胞 髓磷脂碱性蛋白 大鼠 

分 类 号：R595.1[医药卫生—内科学] R747.9[医药卫生—临床医学]