人脐带间充质干细胞移植治疗心肌肥厚模型小鼠  

Human umbilical cord mesenchymal stem cell transplantation for myocardial hypertrophy in mice

作　　者：颉丽英 刘司麒 吴明芮 高尔和 韩之波 左琳[1] Xie Liying;Liu Siqi;Wu Mingrui;Gao Erhe;Han Zhibo;Zuo Lin(Key Laboratory of Cellular Physiology,Ministry of Education,and the Department of Physiology,Shanxi Medical University,Taiyuan 030001,Shanxi Province,China;Center for Translational Medicine,Temple University School of Medicine,Philadelphia,PA,USA;Hematology Hospital/Institute of Hematology,Chinese Academy of Medical Sciences,Tianjin Oncay Cell Genetic Engineering Co.,Ltd./National Engineering Research Center for Cell Products,Tianjin 300041,China)

机构地区：[1]山西医科大学生理学系,教育部细胞生理学重点实验室,山西省太原市030001 [2]天普大学医学院转化医学中心,美国宾夕法尼亚州费城 [3]中国医学科学院血液病医院/中国医学科学院血液学研究所、天津昂赛细胞基因工程有限公司/细胞产品国家工程研究中心,天津市300041

出　　处：《中国组织工程研究》2022年第30期4826-4833,共8页Chinese Journal of Tissue Engineering Research

基　　金：山西省青年基金资助项目(201601D202106),项目负责人:左琳;山西省回国留学人员科研基金资助项目(2014-036),项目负责人:左琳;山西“1331工程”重点学科建设(1331KSC,XK201708),项目参与人:左琳。

摘　　要：背景:左心室肥厚作为高血压病常见的靶器官损害,是心脑血管疾病高发病率和死亡率的独立危险因素。间充质干细胞可以促进组织修复,调节免疫反应,可作为治疗心肌肥厚的种子细胞。目的:观察人脐带间充质干细胞静脉注射治疗小鼠心肌肥厚的疗效。方法:①将C57BL/6J小鼠随机分为假手术组、人脐带间充质干细胞组、PBS组,其中人脐带间充质干细胞组和PBS组采用主动脉弓缩窄术构建C57BL/6J小鼠心肌肥厚模型,人脐带间充质干细胞组小鼠术后第5周开始,尾静脉注射细胞混悬液,每周1次,连续4周,PBS组小鼠尾静脉注射等量PBS,术后6-11周采用超声心动图观察各组小鼠心功能,术后9,10,11周采用苏木精-伊红染色、Western blot检测心肌细胞肥大程度;②体外实验中,通过异丙肾上腺素诱导H9c2细胞肥大,然后将人脐带间充质干细胞与肥大的H9c2细胞共培养于Transwell^(TM)小室中,共培养48 h后F-actin染色分析H9c2细胞横截面积,qRT-PCR检测H9c2细胞内心房钠尿肽、脑钠尿肽和β-肌球蛋白重链mRNA表达量。结果与结论:①超声心动图结果证实:人脐带间充质干细胞组小鼠左心室射血分数、左室短轴缩短率明显高于同期PBS组(P<0.05);②苏木精-伊红染色结果发现,相比于PBS组,人脐带间充质干细胞组心肌纤维排列整齐,心肌细胞横截面积明显减小(P<0.05);③Western blot实验结果证明,人脐带间充质干细胞组小鼠心肌组织中心房钠尿肽、脑钠尿肽和β-肌球蛋白重链蛋白水平明显低于同期PBS组(P<0.01);④与异丙肾上腺素组相比,共培养后的H9c2细胞面积明显减小(P<0.01),同时,H9c2细胞中心房钠尿肽、脑钠尿肽和β-肌球蛋白重链mRNA水平明显减少(P<0.05);⑤结果表明,人脐带间充质干细胞可通过抑制细胞内肥大相关基因的表达,在一定程度上缓解压力负荷型心肌肥厚的发生,改善小鼠心功能。BACKGROUND:As a common target organ damage in hypertension,left ventricular hypertrophy is an independent risk factor for high morbidity and mortality of cardiovascular and cerebrovascular diseases.Mesenchymal stem cells can promote tissue repair,regulate immune response,and can be used as seed cells for the treatment of cardiac hypertrophy.OBJECTIVE:To observe the efficacy of intravenous injection of human umbilical cord mesenchymal stem cells in the treatment of myocardial hypertrophy in mice.METHODS:(1)C57BL/6J mice were randomly divided into a sham operation group,a human umbilical cord mesenchymal stem cell group,and a PBS group.Among them,transverse aortic constriction was used to construct a C57BL/6J mouse cardiac hypertrophy model in the human umbilical cord mesenchymal stem cell group and the PBS group.The mice in the human umbilical cord mesenchymal stem cell group were injected with the cell suspension into the tail vein once a week for 4 weeks starting from the 5th week after the operation.The mice in the PBS group were injected with the same amount of PBS in the tail vein.At 6-11 weeks after operation,echocardiography was used to observe the cardiac function of the mice in each group.Hematoxylin-eosin staining and western blot assay were used to detect the degree of cardiomyocyte hypertrophy at 9,10,and 11 weeks after surgery.(2)In vitro experiment,H9c2 cells hypertrophy was induced by isoproterenol.Afterwards,the human umbilical cord mesenchymal stem cells and hypertrophic H9c2 cells were co-cultured in TranswellTM chamber for 48 hours.F-actin staining was used to analyze H9c2 cells area.qRT-PCR was utilized to determine mRNA expression of atrial natriuretic peptide,brain natriuretic peptide,andβ-myosin heavy chain in H9c2 cells.RESULTS AND CONCLUSION:(1)The results of echocardiography confirmed that the left ventricular ejection fraction and left ventricular fraction shortening of mouse heart in the human umbilical cord mesenchymal stem cell group were significantly higher than those in the PBS gr

关 键 词：人脐带间充质干细胞 小鼠 主动脉弓缩窄术 细胞肥大 细胞共培养 心房利钠肽 脑利钠肽 肌球蛋白重链 

分 类 号：R459.9[医药卫生—治疗学] R331[医药卫生—临床医学]