miRNA-125a在脑卒中的作用及其机制研究

The Role and Mechanism of miRNA-125a in Stroke

目的:探究miR-125a在脑卒中的作用及其分子机制,为miR-125a在脑缺血治疗中的应用提供理论基础。方法:收集正常和脑卒中患者的血浆,Realtime PCR检测miR-125a的表达水平,ROC曲线分析敏感性和特异性。Person相关性分析miR-125a表达水平与患者NIHSS评分和脑梗死体积的相关性。Realtime PCR检测MCAO模型大鼠脑皮质和血浆中miR-125a表达水平,并进行相关性分析。9.4T磁共振测量MCAO模型大鼠脑梗死体积。Realtime PCR和Western Blot法检测AQP4 m RNA和蛋白的表达水平。结果:研究一共收集到70例正常和50例缺血性脑卒中患者的血浆样本,两组在性别(χ^(2)=1.469,P=0.225)和年龄(Z=-0.208,P=0.835)上无差异。缺血性脑卒中患者血浆中miR-125a表达水平高于正常组(Z=-7.01,P=0.000)。预后良好患者血浆中miR-125a表达水平低于预后不良患者(Z=-2.183,P=0.029)。血浆中miR-125a表达水平在区分正常者和缺血性脑卒中患者的敏感性和特异性较好(ROC=0.876,P=0.000)。血浆中miR-125a表达水平与患者NIHSS评分正相关(r=0.303,P=0.032),和脑梗死体积正相关性(r=0.399,P=0.004)。MCAO大鼠脑皮质中miR-125a表达水平升高(t=8.918,P=0.000),血浆中miR-125a表达水平升高(t=4.928,P=0.000),脑皮质与血浆中miR-125a表达水平呈正相关(r=0.823,P=0.044)。第3天时,sh-miR-125a组脑梗死体积小于NC组。sh-miR-125a组AQP4 m RNA和蛋白表达水平小于NC组。结论:血浆中miR-125a表达水平在区分正常和脑卒中患者具有较好的敏感性和特异性,抑制miR-125a表达会通过降低AQP4的表达,减少脑梗死体积,为miR-125a在诊断和治疗脑缺血中的应用提供了理论基础。

Objective:To explore the role and mechanism of miR-125 a in stroke,and provide a theoretical basis for the application of miR-125 a in the treatment of cerebral ischemia.Methods:Collected the plasma of normal and stroke patients,detected the expression of miR-125 a by Realtime PCR,and analyze the sensitivity and specificity of ROC curve.Person correlation analyze the correlation between the miR-125 a level and patient’s NIHSS score,and cerebral infarction volume.Realtime PCR detected the expression of miR-125 a in the cerebral cortex and plasma of MCAO model rats,and analyze the correlation.9.4 T magnetic resonance measurement of cerebral infarction volume in MCAO model rats.Realtime PCR and Western Blot methods were used to detect the expression of AQP4 m RNA and protein.Results:70 normal and 50 ischemic stroke patients’plasma samples were collected in this study.There was no difference in gender(χ^(2)=1.469,P=0.225)and age(Z=-0.208,P=0.835)in the two groups.The expression of miR-125 a in the plasma of ischemic stroke patients was higher than that in the normal group(Z=-7.01,P=0.000).The expression of miR-125 a in the plasma of patients with good prognosis was lower than that of patients with poor prognosis(Z=-2.183,P=0.029).The expression of miR-125 a in plasma has good sensitivity and specificity in distinguishing normal patients from ischemic stroke patients(ROC=0.876,P=0.000).The expression of miR-125 a in plasma was positively correlated with the patient’s NIHSS score(r=0.303,P=0.032),and positively correlated with cerebral infarction volume(r=0.399,P=0.004).The expression of miR-125 a in the cerebral cortex of MCAO rats increased(t=8.918,P=0.000),the expression level of miR-125 a in the plasma increased(t=4.928,P=0.000),and the level of miR-125 a in the cerebral cortex and plasma was increased and they were positively correlated(r=0.823,P=0.044).On the 3 rd day,the cerebral infarction volume of the miR-125 a knockdown group was smaller than the blank control group.The m RNA and protein expression of AQP4 in the miR-125 a knockdown group were lower than the blank control group.Conclusion:The expression of miR-125 a in plasma has good sensitivity and specificity in distinguishing between normal and stroke patients.Inhibition of miR-125 a expression will reduce the expression of AQP4 and the volume of cerebral infarction,which provides a theoretical basis for miR-125 a in diagnosis and treatment of cerebral ischemia.