HGF/c-Met轴介导EPCs治疗大鼠HPAH的初步实验研究  

Effects of HGF/c-Met-mediated endothelial progenitor cells on pulmonary artery pressure and pulmonary vascular remodeling in rats with hypoxic pulmonary hypertension: A preliminary experimental study

作　　者：白辉科 秦廷江 杜明成[1] 刘盈贝 何瑞靖 栾果 普金海 安永[1] Bai Huike;Qin Tingjiang;Du Mingcheng;Liu Yingbei;He Ruijing;Luan Guo;Pu Jinhai;An Yong(Department of Cardiovascular and Thoracic Surgery,Children's Hospital of Chongqing Medical University\Key Laboratory of Pediatrics in Chongq ing,Ministry of Education Key Laboratory of Child Development and Disorders,China International Science and Technology Cooperation Base of Child Development and Critical Disorders)

机构地区：[1]重庆医科大学附属儿童医院心胸外科、儿童发育疾病研究省部共建教育部重点实验室、儿科学重庆市重点实验室、重庆市儿童发育重大疾病诊治与预防国际科技合作基地

出　　处：《重庆医科大学学报》2019年第11期1506-1512,共7页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目（编号：81370432）;重庆市自然科学基金资助项目（编号：cstc2016jcyjA0384）;重庆市教委科学基金资助项目（编号：KJ1500238）;重庆市高端医学后备人才计划资助项目;中华医学会心胸外科分会厄尔巴肯基金资助项目

摘　　要：目的:探讨肝细胞生长因子(hepatocyte growth factor,HGF)/c-Met轴介导内皮祖细胞(endothelial progenitor cells,EPCs)移植入低氧性肺动脉高压(hypoxic pulmonary artery hypertension,HPAH)大鼠对大鼠的肺动脉压力和肺血管重构的影响。方法:构建HPAH模型大鼠;大鼠骨髓原代EPCs提取、培养和鉴定;构建c-Met腺病毒并转染EPCs;将大鼠分为c-Met-EPCs细胞组、EPCs细胞组、肺动脉高压组和正常组,每组10只。前2组大鼠通过尾静脉分别输注c-Met-EPCs细胞和EPCs细胞,后2组大鼠输注生理盐水;饲养一周后检测平均肺动脉压力(mean pulmonary arterial pressure,m PAP)和右心室肥大指数(right ventricle hypertrophy index,RVHI);透射电镜检测肺动脉超微结构改变,肺组织病理切片观察肺小动脉改变;ELISA法测定血清内皮素-1(endothelin-1,ET-1)含量,Griess法测定血清一氧化氮(nitric oxide,NO)含量。结果:c-Met-EPCs细胞组、EPCs细胞组、肺动脉高压组和正常组4组大鼠的mPAP值分别为(23.17±3.07)、(30.85±3.15)、(33.55±5.47)、(20.30±1.99)mmHg;RVHI值分别为(32.48±2.38)%、(37.54±4.42)%、(38.53±2.81)%、(26.05±3.05)%;血清ET-1值分别为(69.45±6.32)、(95.76±7.31)、(99.14±8.39)、(62.35±6.06)ng/L;血清NO值分别为(99.63±11.40)、(56.07±3.32)、(48.83±6.56)、(125.50±12.26)μmol/L。EPCs细胞组、肺动脉高压组大鼠的mPAP、RVHI和血清ET-1含量明显高于正常组(P<0.05);c-Met-EPCs细胞组较EPCs细胞组、肺动脉高压组明显降低(P<0.05)。EPCs细胞组、肺动脉高压组大鼠的血清NO含量明显低于正常组(P<0.05);c-Met-EPCs细胞组较EPCs细胞组、肺动脉高压组明显升高(P<0.05)。透射电镜显示EPCs细胞组、肺动脉高压组肺动脉中膜平滑肌细胞线粒体和内质网肿胀、内皮细胞中断不连续,而c-Met-EPCs细胞肺动脉中膜平滑肌细胞线粒体和内质网肿胀不显著。肺组织病理切片显示EPCs细胞组、肺动脉高压组肺血管重构明显,而c-Met-EPCs细胞�Objective:To investigate the effects of hepatocyte growth factor(HGF)/c-Met-mediated endothelial progenitor cells(EPCs) on pulmonary artery pressure(PAP)and pulmonary vascular remodeling in rats with hypoxic pulmonary hypertension(HPH).Methods:A rat model of HPH was established.Primary EPCs were isolated from the bone marrow of rats,cultured,and identified.The recombinant adenovirus plasmid carrying the c-Met gene was constructed and transfected into EPCs.The rats were divided into c-Met-EPCs group,EPCs group,pulmonary hypertension group,and normal group,with 10 rats in each group.c-Met-EPCs and EPCs were infused into the tail vein of rats in the c-Met-EPCs group and EPCs group,respectively,while normal saline was infused into the tail vein of rats in the pulmonary hypertension group and normal group.After normal feeding for a week,the mean PAP(mPAP)and right ventricular hypertrophy index(RVHI)were determined.The ultrastructural pathology of the pulmonary artery was observed under a transmission electron microscope,and the pathological sections of lung tissues were prepared to observe the histopathological changes of pulmonary arterioles.ELISA was used to determine the serum level of endothelin-1(ET-1),and the Griess method was used to determine the serum level of nitric oxide(NO).Results:The measured values of mPAP for the c-Met-EPCs group,EPCs group,pulmonary hypertension group,and normal group were(23.17±3.07)mmHg,(30.85±3.15)mmHg,(33.55±5.47)mmHg,and(20.30±1.99)mmHg,respectively;the measured values of RVHI for the four groups were(32.48±2.38)%,(37.54±4.42)%,(38.53±2.81)%,and(26.05±3.05)%,respectively;the serum level of ET-1 for the four groups were(69.45±6.32)ng/L,(95.76±7.31)ng/L,(99.14±8.39)ng/L,and(62.35±6.06)ng/L,respectively;the serum level of NO for the four groups were(99.63±11.40)μmol/L,(56.07±3.32)μmol/L,(48.83±6.56)μmol/L,and(125.50±12.26)μmol/L,respectively.The mPAP,RVHI,and serum ET-1 level in the EPCs group and pulmonary hypertension group were significantly higher than those

关 键 词：低氧性肺动脉高压 肝细胞生长因子 C-MET 内皮祖细胞 

分 类 号：R544.1[医药卫生—心血管疾病] R543.2[医药卫生—内科学]