靶向血管紧张素原RNA干扰对自发性高血压模型大鼠的影响  

Effect of angiotensinogen-targeted RNA interference in spontaneously hypertensive rats

作　　者：袁丽粉[1] 曹爽[2] 孙婷[3] Yuan Lifen;Cao Shuang;Sun Ting(Department of Respiratory Medicine and Critical Care Medicine,Shanghai Ninth People’s Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 200011,China;Department of Anesthesiology,Shanghai Ninth People’s Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 200011,China;Department of Cardiovascular Medicine,Shanghai Ninth People’s Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 200011,China)

机构地区：[1]上海交通大学医学院附属第九人民医院呼吸与危重症医学科,上海市200011 [2]上海交通大学医学院附属第九人民医院麻醉科,上海市200011 [3]上海交通大学医学院附属第九人民医院心血管内科,上海市200011

出　　处：《中国组织工程研究》2021年第35期5626-5631,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金面上项目(81770505),项目负责人:孙婷。

摘　　要：背景:前期研究验证了GPE-AGTshRNA纳米复合物在体外对肝细胞不仅有较好的转染效率、较低的细胞毒性,而且在基因和蛋白水平上明显降低了血管紧张素原的表达。目的:观察靶向血管紧张素原RNA干扰对自发性高血压大鼠血压及其主要靶器官心脏的影响,以期寻找一种新的降压策略。方法:实验分为4组:自发性高血压大鼠随机均分为基因治疗组、阴性对照组、空白对照组;同时设立正常血压对照大鼠为WKY组。实验共计9个周期,10 d为1个周期。于每1个周期的第1天经鼠尾静脉注射给药:WKY组注射无菌注射用水500μL,基因治疗组注射GPE-AGT shRNA纳米复合物500μL,阴性对照组注射GPE-NC shRNA纳米复合物500μL,空白对照组注射无菌注射用水500μL;②Real-time PCR法检测肝脏血管紧张素原mRNA的表达,Western blot法检测肝脏血管紧张素原蛋白的表达,ELISA法测定血清血管紧张素原和血管紧张素Ⅱ的水平,尾袖法测量大鼠尾动脉收缩压,心超检测大鼠心室功能,光镜观察心肌组织结构病变。结果与结论:①基因治疗组大鼠肝脏血管紧张素原mRNA和蛋白质表达以及血清血管紧张素原、血管紧张素Ⅱ水平与阴性对照组、空白对照组相比明显下降(P<0.01);②首次注射后第3天,基因治疗组自发性高血压大鼠尾动脉压显著下降(28±4)mmHg,与治疗前比较有显著性差异(P<0.01);③基因治疗组左心室射血分数、左室短轴缩短率明显升高,左室后壁厚度明显下降,而且光镜下心肌细胞肥大明显减轻;④结果说明,靶向血管紧张素原RNA干扰有效下调了肝脏血管紧张素原基因的表达,控制血压的同时显著改善了心肌肥大及心室功能。BACKGROUND:Recent studies have found that GPE-AGTshRNA nanocomposite cannot only have better transfection efficiency and lower cytotoxicity to hepatocytes in vitro,and also significantly reduce the expression of angiotensinogen at the gene and protein levels.OBJECTIVE:To observe the effects of angiotensinogen-targeted RNA interference on blood pressure and cardiac protection in spontaneously hypertensive rats,attempting to find a new antihypertensive strategy.METHODS:Spontaneously hypertensive rats were randomly evenly divided into gene therapy group,negative control group and blank control group,and normal Wistar Kyoto rats(WKY)were set as normal blood pressure control group(WKY group).There were nine sessions in total,with 10 days as one session.On the 1st day of each session,administration in each group was given via the rat tail vein:500μL of sterile water for injection in the WKY group,500μL of GPEAGTshRNA nanocomplex in the gene therapy group,and 500μL of GPE-NC shRNA nanocomplex in the negative control group,and 500μL of sterile water in the blank control group.We used real-time PCR and western blot assay respectively to detect hepatic angiotensinogen mRNA and protein levels.The serum levels of angiotensinogen and angiotensin Ⅱ were measured by ELISA.Systolic blood pressure of the tail artery was measured with tail cuff method.Ventricular function was detected by echocardiography.Myocardial lesion was observed under light microscopy.RESULTS AND CONCLUSION:Angiotensinogen mRNA and protein expression and serum angiotensinogen and angiotensin Ⅱ levels were significantly lower in the gene therapy group than the negative and blank control groups(P<0.01).On the 3rd day after the first injection,tail arterial pressure of rats decreased significantly by(28±4)mmHg in the gene therapy group,which was significantly different from that before treatment(P<0.01).The parameters of left ventricular ejection fraction and left ventricular fractional shortening were significantly increased in the gene therapy group,

关 键 词：高血压 血管紧张素原 RNA干扰 自发性高血压大鼠 基因治疗 心肌肥大 

分 类 号：R446[医药卫生—诊断学] R496[医药卫生—临床医学]