张继厚,陈权,刘美,尚游
ZHANG Jihou,CHEN Quan,LIU Mei,SHAMG You

• 1:锦州医科大学附属第一医院麻醉科
• 2:中国医科大学药学院

摘要(Abstract)：

目的 通过生物信息学分析右美托咪定(dexmedetomidine, DEX)对大鼠心肌miRNA及mRNA表达谱的影响，构建miRNA-mRNA调控网络，探究DEX在大鼠心脏保护中的作用机制。方法 从公共基因芯片数据平台下载经DEX预处理大鼠缺血再灌注损伤心肌的miRNA基因芯片数据集GSE126105和mRNA数据集GSE126104,筛选差异表达miRNA和mRNA。利用miRWalk数据库对差异表达的miRNA靶基因进行预测。用Metascape数据库对预测靶基因进行基因本体论(gene ontology, GO)和基因组百科全书(kyoto encyclopedia of genes and genomes, KEGG)信号通路富集分析，应用Cytoscape对模块中基因的共表达关系可视化并应用cytoHubba插件筛选Hub基因。结果 给药组大鼠心肌较对照组有5个miRNA和165个mRNA差异表达。将miRWalk数据库预测的miRNAs的靶基因和差异表达的mRNA应用韦恩图取交集，得到42个共同基因，基于此结果构建了miRNA-mRNA调控网络，对此网络基因进行功能富集分析，发现细胞周期增殖、蛋白激酶活性等生物过程(biological process, BP)。筛选出前几个的Hub基因包括Ccnd1、Cdk14和Grm5,主要涉及细胞增殖、RNA转录等过程。结论 通过生物信息学分析发现DEX主要通过调控细胞增殖及炎症通路发挥心肌保护作用，构建的miRNA-mRNA调控网络为进一步探究DEX在心脏I/R损伤及在其他疾病中的作用机制提供参考。
Objective To analyze the effects of DEX on rat myocardial miRNA and mRNA expression profiles by using bioinformatics data mining; to construct miRNA-mRNA regulatory network, and to explore the mechanism of dexmedetomidine(DEX) in rat cardioprotection.Methods The miRNA gene chip datasets GSE126105 and mRNA datasets GSE126105 were download from public gene chip data platform(Gene expression omnibus, GEO) for screening differentially expressed miRNA and mRNA.The miRWalk database was used to predict the miRNA target genes.Metascape database was utilized to analyze gene ontology(GO) and KEGG signaling pathway enrichment.Cytoscape was used to visualize the co-expression of genes in the module and predict Hub genes by cytoHubba.Results Compared with the control group, 5 miRNAs and 165 mRNAs were differentially expressed in DEX group.Biological progress of differentially expressed mRNA mainly involves in cell cycle, MAPK pathway, et al.The target genes of miRNAs predicted by miRWalk database and differentially expressed mRNAs were intersected by Venn diagram, and 42 shared genes were obtained, and miRNA-mRNA regulatory network was constructed based on these results.Functional enrichment analysis revealed that biological processes(BP) were mainly involved in cell cycle proliferation and protein kinase activity, etc.The top Hub genes included Ccnd1,Cdk14 and Grm5,which were mainly involved in cell proliferation, RNA transcription and other processes.Conclusion Our study found that DEX plays a protective role in the cardiac muscle mainly by regulating cell proliferation and inflammatory pathways, and the constructed miRNA-mRNA regulatory network provides a reference for further exploring DEX therapeutic role in cardiac I/R damage and other diseases.

关键词(KeyWords)： 右美托咪定;缺血再灌注;miRNA-mRNA网络;生物信息学
dexmedetomidine;ischemia-reperfusion;miRNA-mRNA network;bioinformatics

Abstract:

Keywords:

基金项目(Foundation): 中国博士后科学基金资助项目(2018M641738)

作者(Author): 张继厚,陈权,刘美,尚游
ZHANG Jihou,CHEN Quan,LIU Mei,SHAMG You

DOI: 10.14066/j.cnki.cn21-1349/r.2021.0650

参考文献(References)：

• [1] WU X Y,MIAO L,ZHENG R,et al.Research progress of myocardial ischemia-reperfusion injury [J].The Chinese Journal of Clinical Pharmacology(中国临床药理学杂志),2016(11):1043-1045.
• [2] QIAN Q,LI Y Q.Analysis of the effectiveness of dexmedetomidine nasal drops in the prevention of agitation during recovery from sevoflurane anesthesia in children [J].Journal of Shenyang Pharmaceutical University(沈阳药科大学学报),2021,38(S1):54.
• [3] YOSHIKAWA Y,HIRATA N,KAWAGUCHI R,et al.Dexmedetomidine maintains its direct cardioprotective effect against ischemia/reperfusion injury in hypertensive hypertrophied myocardium[J].Anesth Analg,2018,126(2):443-452.
• [4] CHEN S,HUA F,LU J,et al.Effect of dexmedetomidine on myocardial ischemia-reperfusion injury[J].Int J Clin Exp Med,2015,8(11):21166-21172.
• [5] LIU Y,HUANG D D.MiR-124-3p reversed cisplatin resistance of cervical cancer cells through targeting downregulating FOXC1[J].Journal of Shenyang Pharmaceutical University(沈阳药科大学学报),2022,39(2):180-186.
• [6] OLSON J M,YAN Y,BAI X,et al.Up-regulation of microRNA-21 mediates isoflurane-induced protection of cardiomyocytes[J].Anesthesiology,2015,122(4):795-805.
• [7] TIAN Y K,FU Q,DONG F Q,et al.Up-regulation of microRNA-133a protects against myocardial ischemia/reperfusion injury[J].Chinese Journal of Experimental Surgery(中华实验外科杂志),2012,29(2):270-272.
• [8] IBACACHE M,SANCHEZ G,PEDROZO Z,et al.Dexmedetomidine preconditioning activates pro-survival kinases and attenuates regional ischemia/reperfusion injury in rat heart[J].Biochim Biophys Acta,2012,1822(4):537-545.
• [9] SZKLARCZYK D,MORRIS J H,COOK H,et al.The STRING database in 2017:quality-controlled protein-protein association networks,made broadly accessible[J].Nucleic Acids Res,2017,45(D1):362-368.
• [10] DONCHEVA N T,MORRIS J H,GORODKIN J,et al.Cytoscape StringAppstringApp:network network analysis and visualization of proteomics datadata[J].J Proteome Res,2019,18(2):623-632.
• [11] ZHOU Q L,TENG F,ZHANG Y S,et al.FPR1 gene silencing suppresses cardiomyocyte apoptosis and ventricular remodeling in rats with ischemia/reperfusion injury through the inhibition of MAPK signaling pathway[J].Exp Cell Res,2018,370(2):506-518.
• [12] LAVON H,MATZNER P,BENBENISHTY A,et al.Dexmedetomidine promotes metastasis in rodent models of breast,lung,and colon cancers[J].Br J Anaesth,2018,120(1):188-196.
• [13] CHEN H Y,LI G H,TAN G C,et al.Dexmedetomidine enhances hypoxia-induced cancer cell progression[J].Exp Ther Med,2019,18(6):4820-4828.
• [14] CHEN X L,ZHENG D G,XU Q W,et al.Effects of south semen lepidii extraction solution on myocardial autophagy and MAPK/ERK1/2 pathway in rats with myocardial ischemia/reperfusion[J].Journal of Emergency in Traditional Chinese Medicine(中国中医急症),2019,28(8):1349-1353.
• [15] ZHENG A,CAO L,QIN S,et al.Exenatide regulates substrate preferences through the p38gamma MAPK pathway after ischaemia/reperfusion injury in a rat heart[J].Heart Lung Circ,2017,26(4):404-412.
• [16] GONG X Y,ZHANG Y.Protective effect of miR-20a against hypoxia/reoxygenation treatment on cardiomyocytes cell viability and cell apoptosis by targeting TLR4 and inhibiting p38 MAPK/JNK signaling[J].In vitro Cell Dev Biol Anim,2019,55(10):793-800.
• [17] IBACACHE M,SANCHEZ G,PEDROZO Z,et al.Dexmedetomidine preconditioning activates pro-survival kinases and attenuates regional ischemia/reperfusion injury in rat heart[J].Biochim Biophys Acta,2012,1822(4):537-545.
• [18] SHARIATI M,NIKNAFS B,SEGHINSARA A M,et al.Administration of dexamethasone disrupts endometrial receptivity by alteration of expression of miRNA 223,200a,LIF,Muc1,SGK1,and ENaC via the ERK1/2-mTOR pathway[J].J Cell Physiol,2019,234(11):19629-19639.
• [19] LV Z C,CAO X Y,GUO Y X,et al.MiR-137-5p alleviates inflammation by upregulating IL-10R1 expression in rats with spinal cord injury[J].Eur Rev Med Pharmacol Sci,2019,23(11):4551-4557.
• [20] ZHANG L,YANG F,YAN Q.Candesartan ameliorates vascular smooth muscle cell proliferation via regulating miR-301b/STAT3 axis[J].Hum Cell,2020,33(3):528-536.
• [21] SHI Y,LI K,XU K,et al.MiR-155-5p accelerates cerebral ischemia-reperfusion injury via targeting DUSP14 by regulating NF-kappaB and MAPKs signaling pathways[J].Eur Rev Med Pharmacol Sci,2020,24(3):1408-1419.
• [22] CHEN J G,XU X M,JI H,et al.Inhibiting miR-155 protects against myocardial ischemia/reperfusion injury via targeted regulation of HIF-1alpha in rats[J].Iran J Basic Med Sci,2019,22(9):1050-1058.
• [23] PEDRETTI S,BRULHART-MEYNET M C,MONTECUCCO F,et al.HDL protects against myocardial ischemia reperfusion injury via miR-34b and miR-337 expression which requires STAT3[J].PLoS One,2019,14(6):e218432.
• [24] HUANG R,MA J,NIU B,et al.MiR-34b protects against focal cerebral ischemia-reperfusion (I/R) injury in rat by targeting keap1[J].J Stroke Cerebrovasc Dis,2019,28(1):1-9.
• [25] LI N,ZENG J,SUN F,et al.p27 Inhibits CDK6/CCND1 complex formation resulting in cell cycle arrest and inhibition of cell proliferation[J].Cell Cycle,2018,17(19/20):2335-2348.
• [26] MARGARIT E,ARMAS P,GARCIA S N,et al.CNBP modulates the transcription of Wnt signaling pathway components[J].Biochim Biophys Acta,2014,1839(11):1151-1160.
• [27] ZHANG Z,HU F,LIU Y,et al.Activation of type 5 metabotropic glutamate receptor promotes the proliferation of rat retinal progenitor cell via activation of the PI-3-K and MAPK signaling pathways[J].Neuroscience,2016,322:138-151.
• [28] YU L,ZHANG W,HUANG C,et al.FoxO4 promotes myocardial ischemia-reperfusion injury:the role of oxidative stress-induced apoptosis[J].Am J Transl Res,2018,10(9):2890-2900.
• [29] JIN Y Y,ZHAI C L,SHEN Z,et al.Protective effect of sodium tanshinone iiab against rat myocardial ischemia-reperfusion injury[J].Chinese Archives of Traditional Chinese Medicine(中华中医药学刊),2014,32(6):1414-1416.