脑动脉瘤破裂出血后脑脊液中miRNA-3177-3p与脑血管痉挛的发病关系及临床意义  

Association of mi RNA-3177-3p in cerebrospinal fluid with cerebral vasospasm after aneurysmal subarachnoid hemorrhage and its clinical significance

作　　者：翁传波 江涛[1,2,3] 张洋 刘师林 江录伟 王绪扣 WENGChuanbo;JIANGTao;ZHANGYang;LIUShilin;JIANGLuwei;WANGXukou(Department of Neurosurgery,The First Affiliated Hospital of Anhui Medical University,Hefei,Anhui 230041,China;Department of Neurosurgery,Anhui Public Health Clinical Center,Hefei,Anhui 230041,China;Anhui Provincial Institute of Translational Medicine,Hefei,Anhui 230031,China;Department of Neurosurgery,The Second Affiliated Hospital of Anhui Medical University,Hefei,Anhui 230031,China)

机构地区：[1]安徽医科大学第一附属医院神经外科,安徽合肥230041 [2]安徽省公共卫生临床中心神经外科,安徽合肥230041 [3]安徽省转化医学研究院,安徽合肥230031 [4]安徽医科大学第二附属医院神经外科,安徽合肥230031

出　　处：《国际神经病学神经外科学杂志》2023年第3期6-11,共6页Journal of International Neurology and Neurosurgery

基　　金：安徽医科大学校科研基金项目(2019xkj067);安徽省转化医学研究院科研基金项目(2021zhyx-C72)。

摘　　要：目的 测定动脉瘤性蛛网膜下腔出血(aSAH)后脑血管痉挛(CV)患者与脑血管未痉挛患者脑脊液中miRNA-3177-3p的表达情况,探讨其与CV的发病关系及临床意义。方法 将29例aSAH患者分成脑血管痉挛组(CV组)与脑血管未痉挛组(nonCV组)。比较两组患者的一般临床资料;通过生物信息数据分析确定miRNA-3177-3p作为研究对象,测定两组患者脑脊液中miRNA-3177-3p实际表达情况并比较差异;预测miRNA-3177-3p下游靶基因ATP1B3,构建ATP1B3质粒,双萤光素酶报告实验验证miRNA-3177-3p与ATP1B3靶向关系;最后,ELISA法测定两组患者脑脊液中ATP1B3转录产物ATP1β3含量并比较差异。结果 CV组患者Hunt评级高于non-CV组(P<0.05);CV组脑脊液中miRNA-3177-3p表达丰度为(0.873±0.044),与non-CV组(0.908±0.034)比较,差异有统计学意义(t=2.157,P=0.040);双萤光素酶报告实验结果显示,与miRNA-3177-3p作用后,ATP1B3-3’UTR区相较对照组活性降低15%(P<0.05);将ATP1B3-3’UTR区进行突变,ATP1B3-3’UTR区相较突变前活性上升14%(P<0.05);ELISA法证实表达产物ATP1β3在CV组脑脊液中(1.776±0.013)ng/mL高于non-CV组(1.722±0.016)ng/mL(t=7.778,P=0.000)。结论 CV患者脑脊液中miRNA-3177-3p表达下调,ATP1B3高表达可能参与了CV的发病过程,该机制可能在预测与治疗CV上具有一定临床意义。Objective To investigate the association of miRNA-3177-3p with cerebral vasospasm(CV) and its clinical significance by measuring the expression of miRNA-3177-3p in cerebrospinal fluid(CSF) in patients with or without CV after aneurysmal subarachnoid hemorrhage(aSAH).Methods A total of 29 aSAH patients were divided into CV group and non-CV group.General clinical data were compared between the two groups;miRNA-3177-3p was determined as the target miRNA after integrated bioinformatics analysis,and the expression of miRNA-3177-3p in CSF was measured and compared between the two groups;after ATP1B3 was predicted as the downstream target gene of miRNA-3177-3p,ATP1B3plasmids were constructed and the dual-luciferase reporter gene assay was used to verify the targeting relationship between miRNA-3177-3p and ATP1B3;finally,ELISA was used to measure the content of ATP1β3,a transcription product of ATP1B3,in CSF,which was then compared between the two groups.Results The CV group had a significantly higher Hunt grade than the non-CV group(P<0.05).There was a significant difference in the expression level of miRNA-3177-3p in CSF between the CV group and the non-CV group(0.873±0.044 vs 0.908±0.034,t = 2.157,P = 0.040).The dual-luciferase reporter gene assay showed that after interaction with miRNA-3177-3p,the activity of ATP1B3-3'UTR region was reduced by 15% compared with the control group(P<0.05),while the activity of ATP1B3-3'UTR region was increased by 14% after mutation(P<0.05).ELISA showed that the CV group had a significantly higher level of ATP1β3 in CSF than the non-CV group(1.776±0.013 ng/mL vs 1.722±0.016 ng/mL,t = 7.778,P = 0.000).Conclusion The expression level of miRNA-3177-3p is downregulated in CV patients,and the high expression of its target gene ATP1B3 may be involved in the pathogenesis of CV.This mechanism may have a certain clinical significance in the prediction and treatment of CV.

关 键 词：动脉瘤破裂出血 脑血管痉挛 微小核糖核苷酸 脑脊液 

分 类 号：R651.1[医药卫生—外科学]