小鼠Sca-1^+干细胞对心肌梗死模型小鼠的疗效及其基因调控机制  

Curative effect of Sca-1 positive stem cells from mice on myocardial infarction in mouse model and its mechanism in gene regulation

作　　者：马秀霞 李端端[1] 申晶 陈思宇 李燕楠 颉丽英 高尔和 左琳[1] MA Xiu-xia;LI Duan-duan;SHEN Jing;CHEN Si-yu;LI Yan-nan;XIE Li-ying;GAO Er-he;ZUO Lin(Key Laboratory of Cellular Physiology,Ministry of Education,Department of Physiology,College of Basic Medicine,Shanxi Medical University,Taiyuan 030001,Shanxci Province,China)

机构地区：[1]山西医科大学细胞生理学教育部重点实验室基础医学院生理学系,山西太原030001 [2]天普大学医学院转化医学中心,美国费城19140

出　　处：《中国生物制品学杂志》2020年第4期382-389,共8页Chinese Journal of Biologicals

基　　金：山西省青年科技研究基金(201601D202106);山西省回国留学人员科研基金项目(2014-036)。

摘　　要：目的探讨小鼠干细胞抗原-1阳性(stem cell antigen-1-positive,Sca-1^+)干细胞(stem cells,SCs)对心肌梗死(myocardial infarction,MI)模型小鼠的疗效及其基因调控机制。方法采用免疫磁珠分选法急性分离2日龄及3、6、9、12月龄小鼠心脏Sca-1^+SCs;通过结扎8周龄小鼠心脏冠状动脉左主降支,建立小鼠急性MI模型,于心梗边缘区分别注射PBS和Sca-1^+SCs悬液。术后行超声心动图测定心功能,Masson三色染色、心重/体重比(HW/BW)用于评价各组Sca-1^+SCs对心脏结构的影响。将各组小鼠心脏Sca-1^+SCs进行全基因组测序,通过STEM软件对转录组基因表达水平进行聚类及GO富集分析,筛选出乳鼠和成年鼠差异表达的基因类型,并采用qRT-PCR法进一步验证目的基因的表达情况。结果 MI术后,与PBS组比较,乳鼠心脏Sca-1^+SCs治疗组小鼠左室射血分数(left ventricular ejection fraction,LVEF)和左室短轴缩短率(left ventricular fraction shortening,LVFS)均明显升高(P <0. 05),HW/BW明显降低(P <0. 05),左室舒张末内径(left ventricular diastolic internal diameter,LVIDd)明显减小(P <0. 05);与成年鼠比较,乳鼠心脏Sca-1^+SCs移植治疗可降低心肌梗死面积,缓解心腔扩大。在乳鼠心脏Sca-1^+SCs中高表达,而成年鼠表达降低的基因共185个,regulation of developmental process条目上富集基因有38个,其中肌球蛋白重链6(myosin heavy chain6 cardiac muscle alpha,Myh6)、胰岛素样生长因子2(insulin-like growth factor 2,Igf2)、成骨细胞特异性因子(periostin,Postn)、神经元再生相关蛋白(neuronal regeneration related protein,Nrep)基因在乳鼠心脏Sca-1^+SCs中具有较高转录水平。与乳鼠比较,成年小鼠心脏Sca-1^+SCs中Myh6、Igf2、Postn、Nrep基因m RNA的表达水平均显著下调(P <0. 05)。结论乳鼠心脏Sca-1^+SCs移植对MI的疗效优于成年小鼠,其原因可能是乳鼠心脏Sca-1^+SCs中Myh6、Igf2、Postn、Nrep等基因的表达水平高于成年小鼠。Objective To investigate the curative effect of stem cell antigen-1 positive(Sca-1^+)stem cells(SCs)from mice on myocardial infarction(MI)in mouse model as well as its mechanism in gene regulation. Methods Sca-1^+SCs were isolated from the hearts of mice at ages of 2 days as well as 3,6,9 and 12 months by immunomagnetic separation. An acute MI model was established with C57 BL/6 J mice by ligation of left main descending branch of coronary artery of mice at age of8 weeks. PBS and Sca-1^+SCs suspension were injected in the border zone of infarcted myocardium respectively. Cardiac function was detected by echocardiography after MI. Masson trichrome-staining and heart weight/body weight(HW/BW)ratio were used to evaluate the effect of Sca-1^+SCs on cardiac structure after MI in various groups. The whole genomes of Sca-1^+SCs in various groups were sequenced,and the differentially expressed genes in suckling and adult mice were subjected to clustering analysis with STEM software and Gene Ontology(GO)analysis. Moreover,the expression levels of target genes was determined by qRT-PCR. Results After MI,compared with those in PBS group at the same time point,the left ventricle ejection fraction(LVEF) and left ventricle fraction shortening(LVFS) in Sca-1^+SCs group increased significantly(P < 0. 05),while the HW/BW and left ventricle diastolic internal diameter(LVID,d)decreased significantly(each P < 0. 05). The treatment by transplantation with Sca-1^+SCs decreased the infarct size and relieved the enlargement of cardiac chambers in suckling mice as compared with those in adult mice. A total of 185 genes were highly expressed in the Sca-1^+SCs of suckling mice,of which the expression level decreased in adult mice. Thirty-eight genes were enriched in the GO term of the‘regulation of developmental process’,among which Myh6,Igf2,Postn and Nrep genes showed higher transcription levels in cardiac Sca-1^+SCs of suckling mice. Compared with those in suckling mice,the expressions of Myh6,Igf2,Postn and Nrep mRNAs in cardiac Sc

关 键 词：心肌梗死 小鼠 干细胞抗原-1 干细胞 心功能 基因调控 

分 类 号：R331[医药卫生—人体生理学]