microRNA作为高血压病治疗靶点的初步实验研究  

Preliminary experimental study of microRNA as a therapeutic target in hypertension

作　　者：王艺璇[1] 刘媛圆[1] 李明瑛[1] 刘秀兰[1] 高飞[1] 杨新春[1] 蔡军[1] 

机构地区：[1]首都医科大学附属北京朝阳医院,北京100020

出　　处：《山东医药》2012年第4期11-13,16,I0001,I0002,共6页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81170244;81050012)

摘　　要：目的比较高血压病患者和健康人外周血单个核细胞miRNAs的表达谱,并研究miR-126作为基因治疗靶点的可行性。方法运用Exiqon公司的miRCURYTM miRNA芯片检测高血压病患者和健康人miRNAs的表达谱。构建miR-126基因敲减慢病毒载体及干扰对照慢病毒载体。自发性高血压大鼠分为高剂量组、低剂量组、干扰对照组、空白对照组,经尾静脉注射慢病毒载体,应用尾袖法每周采集收缩压、舒张压、平均压、心率,连续观察8周后留取心、肾、脑、肝、肺组织标本进行冰冻切片,并在荧光显微镜下观察慢病毒载体在体内的分布情况;计算心体比值;应用天狼星红苦味酸染色及HE染色,观察高血压靶器官损害情况;应用ELISA法测定大鼠血清一氧化氮含量。结果高血压病患者和健康人外周血单个核细胞miRNAs表达谱存在显著差异;miR-126基因敲减慢病毒载体构建成功,注射后,血压、心率、心体比值、组织形态、一氧化氮各组间比较无统计学意义(P均>0.05)。结论成功构建miR-126基因敲减慢病毒载体,miR-126作为基因治疗靶点治疗原发性高血压无明显降压效果,对靶器官无明显保护作用。Objective To detect differential microRNA expression in hypertensive and normotensive patients and to investigate miR-126 as a therapeutic target in primary hypertension using an animal model.Methods Differential microRNA expression patterns in patients with essential hypertension vs.healthy controls were investigated using miRCURYTM miRNA microarray technology from Exiqon.A lentiviral vector based on the miR-126 gene knock-down viral vector and a control lentiviral vector were constructed.Spontaneously hypertensive rats were divided randomly into 4 groups to receive miRNAs.The lentiviral vectors were injected weekly into the tail veins.Systolic blood pressure,diastolic blood pressure,mean arterial pressure,and heart rate were measured weekly.After 8 weeks of virus administration,the animals were sacrificed and frozen sections were prepared from the hearts,kidneys,brains,livers,and lung tissues.The distribution of lentiviral vectors in the tissues was observed by fluorescence microscope.Picric acid,sirius red,and HE staining were used to detect target organ damage,especially in the heart and kidney.ELISA was used to determine the serum nitric oxide content.Results The microRNA expression profiles differed in patients with essential hypertension compared with healthy donors.Heart rate,blood pressure,and the heart-to-body weight ratio（mg/g） did not differ significantly in the four groups after lentiviral vector injection.Similarly,we observed no differences in organ tissue morphology in the four groups,and there were no significant differences in the serum NO levels in the four groups.Conclusion Knocking-down miR-126 gene expression had no significant antihypertensive effect,although the target organs were not damaged by the treatment.

关 键 词：microRNA-126 原发性高血压 基因治疗 

分 类 号：R544.1[医药卫生—心血管疾病]