miR-138通过靶向LCN2在七氟醚诱导的大鼠认知障碍中的作用及机制  

The role and mechanism of miR-138 in sevoflurane-induced cognitive impairment in rats by targeting LCN2

作　　者：郑育秀 刘丽丽[1] 黄泽波[1] 王涛[1] 刘莉芳 ZHENG Yuxiu;LIU Lili;HUANG Zebo;WANG Tao;LIU Lifang(Department of Anesthesiology,The Second Affiliated Hospital of Hainan Medical College,Haikou 570311,China)

机构地区：[1]海南医学院第二附属医院麻醉科,海南海口570311

出　　处：《西部医学》2022年第10期1442-1449,共8页Medical Journal of West China

基　　金：国家自然科学基金委员会资助项目(81660195);海南省卫生健康行业科研项目(20A200379)。

摘　　要：目的探讨miR-138通过靶向LCN2在七氟醚诱导的大鼠认知障碍中的作用及机制。方法将90只7周龄SD大鼠随机分为对照组(n=20)、七氟醚组(n=20)、七氟醚+NC mimic组(n=20)、七氟醚+miR-138 mimic组(n=20)、七氟醚+miR-138 mimic组+vector组(n=5)、七氟醚+miR-138 mimic+pcDNA-LCN2组(n=5)。构建七氟醚诱导的大鼠认知障碍模型,七氟醚诱导的大鼠分别注射10μL miR-138 mimic、NC mimic慢病毒悬液。采用Morris水迷宫实验分析潜伏期、跨台时间和游泳速度;HE染色检测海马神经元的病理变化;TUNEL染色检测海马神经元凋亡情况;实时定量聚合酶链反应(RT-qPCR)检测miR-138、脂质运载蛋白2(LCN2)的mRNA水平表达;双荧光素酶报告基因系统用于检测miR-138和LCN2之间的关系;Western blotting检测海马组织中LCN2、cleaved caspase-3、cleaved caspase-9、Bax、Bcl-2、p-JAK2和p-STAT3蛋白水平;ELISA实验检测海马组织中IL-6、TNF-α、IL-1β。结果与对照组比较,七氟醚组中miR-138表达水平显著降低(P<0.05);大鼠逃避潜伏期显著增加,穿越平台次数显著减少,海马神经元损伤,海马神经元凋亡数量显著增加,cleaved caspase-3、cleaved caspase-9和Bax蛋白水平显著升高,Bcl-2显著降低,IL-1β、TNF-α、IL-6水平升高,p-JAK2和p-STAT3蛋白水平降低(均P<0.05)。与七氟醚+NC mimic组比较,七氟醚+miR-138 mimic组中大鼠逃避潜伏期减少,穿越平台次数增加,神经元病理损伤减轻,海马神经元凋亡数量显著减少,cleaved caspase-3、cleaved caspase-9和Bax蛋白水平降低,Bcl-2水平升高,IL-1β、TNF-α、IL-6水平降低,p-JAK2和p-STAT3蛋白水平升高(均P<0.05)。双荧光素酶报告基因结果显示LCN2是miR-138的靶基因,过表达LCN2可显著逆转miR-138对七氟醚诱导的认知障碍大鼠凋亡的作用(P<0.05)。结论miR-138靶向LCN2通过JAK2/STAT3通路对七氟醚诱导的认知障碍具有神经保护作用。Objective To investigate the neuroprotective effect of miR-138 on cognitive impairment induced by sevoflurane anesthesia.Methods 7-week-old SD rats were randomly divided into control group,sevoflurane group,sevoflurane+NC mimic group,sevoflurane+miR-138 mimic group,sevoflurane+miR-138 mimic vector group,sevoflurane+miR-138 mimic+pcDNA-LCN2 group.A rat model of cognitive impairment induced by sevoflurane was constructed.The rats induced by sevoflurane were injected with 10μL miR-138 mimic and NC mimic lentivirus suspensions.Morris water maze experiment was used to analyze the incubation period,cross-stage time and swimming speed.HE staining was used to detect the pathological changes of hippocampal neurons.TUNEL staining was used to detect the apoptosis of hippocampal neurons.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect miR-138 and lipocalin 2(LCN2)mRNA level expression.The dual luciferase reporter gene system was used to detect the relationship between miR-138 and LCN2.Western blotting was used to detect LCN2,cleaved caspase-3,cleaved caspase-9,Bax,Bcl-2,p-JAK2 and p-STAT3 protein levels.ELISA test was used to detect IL-6,TNF-α,IL-1βin hippocampus.Results Sevoflurane group compared with control group,miR-138 expression level was significantly reduced(P<0.05).The number of hippocampal neuronal damage and apoptosis were significantly increased,the protein levels of cleaved caspase-3,cleaved caspase-9 and Bax were significantly increased,and the protein levels of Bcl-2 were significantly decreased.IL beta,TNF-1 alpha,IL-6 levels,p-JAK2 and p-STAT3 protein levels(P<0.05).Compared with sevoflurane+miR-138 mimic group,the escape latency of rats in sevoflurane+miR-138 mimic group was decreased,The Times of crossing platform was increased,neuronal pathological damage was attenuated,and the number of hippocampal neuronal apoptosis was significantly decreased.Cleaved caspase 3,cleaved-caspase 9-and Bax protein levels are low,the Bcl-2 levels,IL-1 beta,TNF-alpha,IL-6 levels drop,p-JAK2 and

关 键 词：七氟醚 认知障碍 miR-138 LCN2 JAK2/STAT3通路 

分 类 号：R614[医药卫生—麻醉学]