基于circRNA调控网络探讨七氟烷致老年小鼠认知功能障碍的分子机制研究  

To explore the molecular mechanism of sevoflurane induced cognitive dysfunction in elderly mice based on circRNAregulatorynetwork

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,海口570311

出　　处：《脑与神经疾病杂志》2023年第9期538-544,共7页Journal of Brain and Nervous Diseases

基　　金：海南省卫生健康行业科研项目(21A200267)。

摘　　要：目的 基于circRNA调控网络探究circ0014063在七氟烷致老年小鼠认知功能障碍中的作用及其相关机制。方法 circRNA-seq检测七氟烷麻醉和未处理老年小鼠海马组织,根据关键差异circRNA绘制ceRNA调控网络,并对靶基因进行KEGG分析。将30只老年C57/BL6小鼠随机分为:对照组、模型组和治疗组,每组10只。模型组和治疗组使用七氟烷麻醉,治疗组尾静脉注射si-circ0014063,对照组和模型组注射si-NC。水迷宫实验比较各组小鼠认知功能差异,qRT-PCR检测各组小鼠circ0014063、miR-744-3p、Apoe基因表达,western blot检测各组小鼠Caspase3、Bcl-2蛋白表达,ELISA检测各组小鼠β淀粉样蛋白(Aβ)、β淀粉样物质前体蛋白(APP)、IL-1 β、TNF-α含量,并通过spearman法分析circ0014063与A β、APP、IL-1 β、TNF-α的相关性。结果 与未处理组比较,七氟烷麻醉组有238个circRNA表达降低,241个circRNA表达升高,其中circ0014063和circ 0032434表达显著升高(P <0.05)。circRNA调控网络预测circ0014063靶向4个miRNA,18个mRNA,circRNA0032434靶向4个miRNA,16个mRNA。KEGG分析circ0014063靶基因参与阿尔茨海默病(AD)、坏死性凋亡等通路。与对照组比较,模型组小鼠逃脱时间增加,穿台次数降低(P<0.05),海马组织circ0014063、Apoe、Caspase3表达升高(P<0.05),miR-744-3p、Bcl-2表达降低(P<0.05),Aβ、APP、IL-1 β、TNF-α含量升高(P<0.05)。与模型组比较,治疗组小鼠逃脱时间减少,穿台次数升高(P<0.05),海马组织circ0014063、Apoe、Caspase3表达降低(P <0.05),miR-744-3p、Bcl-2表达升高(P<0.05),Aβ、APP、IL-1 β、TNF-α含量降低(P<0.05)。circ0014063表达与Aβ、APP、IL-1 β、TNF-α含量呈正相关。结论 七氟烷可导致老年小鼠认知功能损伤,circRNA调控网络在其中发挥重要作用。七氟烷可能通过调控circ0014063/miR-744-3p/Apoe信号轴介导海马组织凋亡,最终诱发老年小鼠认知功能障碍。Objective Based on the circRNA regulatory network,the role of circ_0014063 in sevoflurane induced cognitive dysfunction in elderly mice and its related mechanisms were explored.Methods circRNA-seq was used to detect hippocamal tissues of sevoflurane anesthetized and untreated aged mice.The ceRNA regulatory network was mapped according to the key differences of circRNA,and the target genes were analyzed by KEGG.Thirty aged C57/BL6 mice were randomly divided into control group,model group and treatment group,with 10 mice in each group.The model group and the treatment group were anesthetized with sevoflurane,si-circ_0014063 was injected into the tail vein of the treatment group,si-NC was injected into the control group and the model group.Water maze test was performed to compare the dfferences in cognitive function of mice in each group.The expressions of circ_0014063,mir-744-3p and Apoe genes were detected by qRT-PCR,and the expressions of Caspase3 and Bcl-2 proteins were detected by Western blot.The contents of amyloid beta(Aβ),amyloid precursor protein(APP),IL-1βand TNF-αwere detected by ELISA,and the correlation between circ_0014063 and Aβ,APP,IL-1βand TNF-αwas analyzed by spearman method.Results Compared with the untreated group,238 circRNA expressions were decreased and 241 circRNA expressions were increased in sevoflurane anesthesia group,among which circ_0014063 and circ_0032434 expressions were significantly increased(P<0.05).The circRNA regulatory network predicted that circ_0014063 would target 4 miRNAs and 18 mRNAs,and circRNA_0032434 would target 4 miRNAs and 16 mRNAs.KEGG analysis of circ_0014063 target gene involved in Alzheimer's disease(AD),necrotic apoptosis and other pathways.Compared with the control group,the escape time of model group was increased and the number of times of platform crossing was decreased(P<0.05),the expressions of circ_0014063,Apoe and Caspase3 in hippocampus were increased(P<0.05),and the expressions of miR-744-3p and Bcl-2 were decreased(P<0.05).The contents of Aβ,A

关 键 词：环状RNA调控网络 七氟烷 认知功能障碍 老年小鼠 

分 类 号：R741.02[医药卫生—神经病学与精神病学]