Survivin对分流型肺动脉高压大鼠肺动脉平滑肌细胞增殖作用及机制研究  

Effect and mechanism of Survivin on proliferation of pulmonary artery smooth muscle cells in rats with shunt pulmonary hypertension

作　　者：黄静林 劳金泉[2] 庞玉生[1] Huang Jinglin;Lao Jinquan;Pang Yusheng(The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China;LiuzhouWorker’s Hospital,Liuzhou 545005,China)

机构地区：[1]广西医科大学第一附属医院,南宁530021 [2]广西柳州市工人医院,柳州545005

出　　处：《广西医科大学学报》2020年第5期837-842,共6页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81660043);广西自然科学基金资助项目(No.2016GXNSFAA380186)。

摘　　要：目的:通过RNA干扰抑制分流型肺动脉高压大鼠肺动脉平滑肌细胞(PASMCs)中survivin的表达,探索survivin基因对PASMCs增殖作用的影响及其机制。方法:将30只SD大鼠随机分为正常组、假手术组和分流组,正常组大鼠不做处理,假手术组大鼠开腹后予夹闭腹主动脉10 min,分流组用腹主动脉-下腔静脉分流术建立肺动脉高压大鼠模型。术后饲养11周后,通过测定大鼠右心室压力、右心室肥厚指数及肺小动脉苏木精-伊红(HE)染色评估模型建立情况。提取各组大鼠肺动脉进行PASMCs原代培养,并通过siRNA干扰技术将分流组大鼠PASMCs在细胞层面上分为分流组、分流+空载病毒组和分流+慢病毒干扰组。用实时荧光定量聚合酶链反应(qPCR)、Western blot分别检测survivin mRNA及蛋白在5组细胞中的表达;CCK-8法检测5组细胞的增殖水平。酶联免疫吸附测定(ELISA)法检测细胞上清液中caspase-9、caspase-3浓度。结果:分流组大鼠的右心室压力、右心室肥厚指数均较正常组和假手术组明显增高(P<0.05),分流组大鼠肺动脉较正常组和假手术组明显增厚。正常组和假手术组的PASMCs均未见survivin mRNA及蛋白表达,分流组为阳性表达。分流+慢病毒干扰组survivin mRNA及蛋白表达水平较分流组、分流+空载病毒组明显下降(P<0.05),分流组、分流+空载病毒组比较,差异无统计学意义(P>0.05)。CCK-8法分析,分流组PAMSCs在接种48 h及之后较正常组和假手术组吸光度值明显增加(P<0.05),正常组和假手术组在各个时间节点比较,差异无统计学意义(P>0.05),分流+慢病毒干扰组PASMCs在接种48 h及之后吸光度值较分流组、分流+空载病毒组明显下降(P<0.05),分流组和分流+空载病毒组在各个时间节点比较,差异无统计学意义(P>0.05)。ELISA法显示分流+慢病毒干扰组caspase-9、caspase-3浓度较分流组、分流+空载病毒中显著增高(P<0.05),分流组与分流+空载病毒组�Objective:RNA interference was used to inhibit the expression of survivin in pulmonary artery smooth muscle cells(PASMCs)of rats with shunt pulmonary hypertension to explore the effect and mechanismof survivin gene on the proliferation of PASMCs.Methods:Thirty SD rats were randomly divided into normal group,sham operation group,and shunt group.The rats in the normal group received no treatment,rats in the sham operation group wereclippedwith abdominal aortafor 10 minafter laparotomy,rats in theshunt groupwereused abdominal aortic-inferior vena cava shuntto established the rat model of pulmonary hypertension.After 11 weeks of feeding,the establishment of the model was evaluated by measuring theright ventricular pressure,right ventricular hypertrophy index,and pulmonary arteriole HE staining.The PASMCsof rat pulmonary arteries from each group were extracted for primary culture.The PASMCs of shunt group were divided into shunt group,shunt+no-load virus group,and shunt+chronic virus interference groupby siRNA interference technology.Real-time fluorescence quantitative polymerase chain reaction(qPCR)and Western blot were used to detect the expression of survivin mRNA and protein in five groups of cells.CCK-8 assay was used to detect the proliferation level infive groups of cells.The level of caspase-9 and caspase-3 in the supernatant of cells were detected by enzyme-linked immunosorbent assay(ELISA).Results:The right ventricular pressure and right ventricular hypertrophy index in the shunt group were significantly higher than those in the normal group and the sham operation group(P<0.05).The pulmonary artery in the shunt group was significantly thicker than that in the normal group and the sham operation group.There was no expression of suvivin mRNA and protein in PASMCs in normal group and sham operation group,but positive expression in shunt group.The expression levels of suvivin mRNA and protein in shunt+chronic virus interference group were significantly lower than those in shunt group and shunt+no-load virus grou

关 键 词：肺动脉高压 SURVIVIN RNA干扰 增殖 大鼠 

分 类 号：R544.1[医药卫生—心血管疾病]