微小核糖核酸mmu-miR-8114和mmu-miR-3473b在细粒棘球蚴致敏小鼠中的调控作用  

REGULATORY EFFECTS OF MMU-MIR-8114 ANDMMU-MIR-3473B IN ALLERGIC MICE INDUCEDBY ECHINOCOCCOSIS

作　　者：马岩[1] 韩静 房志远 苏比·泰来提 于晓东[1] MA Yan;HAN Jing;FANG Zhi-yuan;SUBI·Tailaiti;YU Xiao-dong(Department of Anesthesiology,First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,Xinjiang,China;First People's Hospital of Urumqi,Urumqi 830002,Xinjiang,China;Department of Anesthesiology,First Dongguan Affiliated Hospital of Guangdong Medical University,Dongguan 523710,Guangdong,China;Graduate School,Xinjiang Medical University,Urumqi 830000,Xinjiang,China)

机构地区：[1]新疆医科大学第一附属医院麻醉科,乌鲁木齐830054 [2]乌鲁木齐市第一人民医院,乌鲁木齐830002 [3]广东医科大学附属东莞第一医院麻醉科,广东东莞523710 [4]新疆医科大学研究生学院,乌鲁木齐830000

出　　处：《寄生虫与医学昆虫学报》2024年第3期145-152,162,共9页Acta Parasitologica et Medica Entomologica Sinica

基　　金：自治区卫生健康委员会青年医学技术人才专项(WJWY-202151);新疆维吾尔自治区围术期器官保护重点实验室(XJDX1411)。

摘　　要：目的探索微小核糖核酸(miRNAs)在通过细粒棘球蚴感染引起的小鼠过敏反应中的调节作用。方法建立小鼠体内棘球蚴病动物模型,收集脾脏单核细胞进行转录组测序。比较过敏性小鼠和非过敏小鼠之间mRNAs和miRNAs的差异表达。使用Targetscan和miRanda软件预测mRNAs和miRNAs之间的靶向调控关系,并通过蛋白质—蛋白质相互作用(PPI)网络分析,识别核心基因,通过富集分析探讨核心基因的生物学作用。最后利用qRT-PCR和Western blot验证关键结果。结果过敏与非过敏小鼠之间存在1642个差异表达的mRNAs。对18个差异表达的miRNAs进行分析,鉴定出60个差异表达的靶标基因,其中在PPI网络中连通度最大的前10个基因被认定为核心基因。富集分析显示核心基因主要参与NF-kappa B信号通路。基于miRNAs与mRNAs之间的负调控作用以及双荧光素酶报告系统发现mmu-miR-8114与Atf3以及mmu-miR-3473b与Myd88具有靶向调控关系,并与NF-kappa B信号通路有关。通过qRT-PCR验证了mmu-miR-8114和mmu-miR-3473b在过敏小鼠中下调表达,Atf3、Myd88、Socs3在过敏小鼠中上调表达。qRT-PCR和Western blot结果发现NF-kappa B信号通路在过敏小鼠中的活性增加。结论本研究揭示了mmu-miR-8114和mmu-miR-3473b通过NF-kappa B信号通路调控导致小鼠过敏反应的重要作用,为理解细粒棘球蚴感染引起的过敏反应机制提供了新的见解。Objective This study aimed to explore the regulatory roles of microRNAs(miRNAs)in mice with allergic reactions induced by Echinococcus granulosus.Methods A mouse model of Echinococcosis was established,and spleen mononuclear cells were collected for transcriptomic sequencing.The differential expressions of mRNAs and miRNAs between allergic and non-allergic mice were compared.The target regulatory relationships between mRNAs and miRNAs were predicted using TargetScan and miRanda softwares,and core genes were identified through protein-protein interaction(PPI)network analysis.The biological functions of these core genes were further explored via enrichment analysis.Finally,key results were validated using qRT-PCR and Western blot.Results A total of 1642 differentially expressed mRNAs were identified between allergic and non-allergic mice.Among the 18 differentially expressed miRNAs,60 target genes were identified,with the top 10 genes having the highest connectivity in the PPI network-deemed as core genes.Enrichment analysis showed that these core genes are primarily involved in the NF-kappa B signaling pathway.Based on the negative regulatory effects between miRNAs and mRNAs and the dual-luciferase reporter system,mmu-miR-8114 was found to target Atf3 and mmu-miR-3473b to target Myd88,both of which are related to the NF-kappa B signaling pathway.qRT-PCR validation showed that mmu-miR-8114 and mmu-miR-3473b were downregulated in allergic mice,while Atf3,Myd88,and Socs3 were upregulated.Increased activity of the NF-kappa B signaling pathway was observed in allergic mice through qRT-PCR and Western blot results.Conclusions This study reveals the significant role of mmu-miR-8114 and mmu-miR-3473b in regulating mouse allergic reactions through the NF-kappa B signaling pathway,offering new insights into the mechanisms behind allergic reactions caused by Echinococcus granulosus infection.

关 键 词：细粒棘球蚴 过敏 微小核糖核酸 NF-kappa B信号通路 

分 类 号：R383.3[医药卫生—医学寄生虫学]