基于转录组学探讨阿比朵尔对HCoV-OC43感染诱导神经营养因子信号通路活化的影响  

Transcriptome-based exploration of the effect of Arbidol on HCoV-OC43 infection-induced activation of the neurotrophin signaling pathway

作　　者：陈琪敏 周红霞 谢佩芳 李润峰[4] 杨子峰[4,5,6,7] 邱敏珊 沈利汉 CHEN Qimin;ZHOU Hongxia;XIE Peifang;LI Runfeng;YANG Zifeng;QIU Minshan;SHEN Lihan(Guangdong Medical University,Zhanjiang 524023,China;Department of Critical Care Medicine,Dongguan Institute of Respiratory and Critical Care Medicine,the 10th Affiliated Hospital of Southern Medical University(Dongguan People’s Hospital),Dongguan 523059;Faculty of Life Science and Technology,Kunming University of Science and Technology,Kunming 650500;State Key Laboratory of Respiratory Disease,National Clinical Research Center for Respiratory Disease,Guangzhou Institute of Respiratory Health,the First Affiliated Hospital of Guangzhou Medical University,Guangzhou 510000;Guangzhou Laboratory,Guangzhou 510000;Guangzhou key laboratory for clinical rapid diagnosis and early warning of infectious diseases,Guangzhou 510000;State Key Laboratory of Quality Research in Chinese Medicine,Macao Institute for Applied Research in Medicine and Health,Macao University of Science and Technology,Macao 519020)

机构地区：[1]广东医科大学,广东湛江524023 [2]南方医科大学第十附属医院(东莞市人民院),东莞市呼吸和重症医学研究所,重症医学科,广东东莞523059 [3]昆明理工大学生命科学与技术学院,昆明650500 [4]广州医科大学附属第一医院广州呼吸健康研究院呼吸疾病国家重点实验室/国家呼吸系统疾病临床医学研究中心,广州510000 [5]广州实验室,广州510000 [6]广州市传染性疾病临床快速诊断与预警重点实验室,广州510000 [7]澳门科技大学中药质量研究国家重点实验室,中国澳门519020

出　　处：《中国比较医学杂志》2023年第6期81-89,共9页Chinese Journal of Comparative Medicine

基　　金：广东省防控新型冠状病毒感染科技攻关应急专项(2020A111129028);东莞市新型冠状病毒肺炎防疫防治技术研究及推广应急攻关专项项目(202071715001114);广东省基础与应用基础研究基金(2020B1515120045);广东省基础与应用基础研究基金(2020A1515110151);东莞市人民医院院内项目-科研培育项目(K202002);东莞市人民医院院内项目-科研培育项目(K202019)。

摘　　要：目的采用转录组学技术分析阿比多尔(arbidol,ARB)对HCoV-OC43感染引起宿主信号通路活化的影响,并探讨ARB抗炎活性与其作用于神经营养因子信号通路的关系。方法用OC43感染HRT-18细胞,并以ARB进行干预。感染96 h后提取总RNA,进行转录组学分析,获得差异表达基因(differentially expressed genes,DEGs),开展基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析,筛选出ARB可能作用的生物学过程及相关信号通路。进一步利用RT-qPCR方法验证ARB干预对神经营养因子信号通路关键分子表达的抑制作用。结果与病毒感染组相比,高、中和低剂量(6、2和0.67μg/mL)ARB干预分别显示出9459、4186和1744个DEGs。GO分析显示,ARB主要干预的生物学过程是共翻译蛋白膜靶向,作用的细胞组成为粘着斑和细胞-底物间连接,影响的分子功能为钙粘蛋白结合。KEGG分析发现,与冠状病毒感染相关的细胞凋亡信号通路和神经营养因子信号通路有明显富集,其中ARB对神经营养因子信号通路中的MAPK、PI3K和NF-κB通路分子有显著抑制作用。RT-qPCR检测显示,ARB对PIK3CA、AKT、TRAF-6、Bax、p38和c-JUN等分子的mRNA表达具有明显抑制作用。结论本研究提示了ARB应用于治疗冠状病毒感染引起神经炎症的可能性。Objective To analyze the influence of arbidol(ARB)on HCoV-OC43-induced activation of host signaling pathways and to explore the correlation between the anti-inflammatory activity of ARB and its effect on the neurotrophin signaling pathway.Methods HRT-18 cells were infected with OC43 and treated with ARB.After 96 hours,total RNA was extracted,transcriptomic analysis was performed to identify differentially expressed genes(DEGs),and Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed to identify potential biological processes and signaling pathways related to ARB treatment.RT-qPCR was used to verify the inhibitory effect of ARB on the expression of important molecules in the neurotrophin pathway.Results ARB treatment at high,medium,and low doses(6,2 and 0.67μg/mL)resulted in 9459,4186,and 1744 DEGs,respectively,compared with the virus-infected control group.GO analysis showed that ARB mainly affected biological processes of cotranslational protein targeting to membrane,cellular components such as focal adhesion and cell-substrate junction,and molecular functions such as cadherin binding.KEGG analysis showed that apoptosis and neurotrophin signaling pathways related to coronavirus infection were significantly enriched,and ARB had a significant inhibitory effect on MAPK,PI3K,and NF-κB in the neurotrophin signaling pathway.RT-qPCR analysis revealed that ARB significantly inhibited mRNA expression of PIK3CA,AKT,TRAF-6,Bax,p38,and c-Jun.Conclusions This study suggests that ARB can be used for treatment of neuroinflammation caused by coronavirus infection.

关 键 词：转录组学 阿比多尔 HCoV-OC43 神经营养因子信号通路 

分 类 号：R-33[医药卫生]