中国全科医学2018，Vol.21Issue(2)：167-172,6.DOI:10.3969/j.issn.1007-9572.2018.02.10
过表达GATA-4小鼠骨髓间充质干细胞修复心肌损伤的效果研究
Preliminary Investigation on Repair of Myocardial Damage with Over-expressed GATA-4 BMSCs
摘要
Abstract
Objective This study explores the mechanism of murine bone marrow mesenchymal stem cells(BMSCs) with over-expressed GATA-4 in repairing myocardial injury.Methods From March 2015 to December 2016,sixty healthy SPF male C57BL/6 mice aged 4 weeks were selected.BMSCs were isolated and the expression rates of CD29,CD44,CD11b and SCA-1 were detected by flow cytometry.Over-expressed GATA-4 BMSCs were synthesized by transfecting the murine BMSCs with GATA-4-carrying slow virus vector GV308,subsequently added the gene initiator doxycycline(DOX).The expression levels of GATA-4 mRNA in the over-expressed GATA-4-BMSCs after transfection 72 h were detected by RT-PCR,and no transfection was used as negative control.The myocardial infarction models of mice were successfully established,BMSCs(n=3),blank vector-BMSCs(n=3),over-expressed GATA-4-BMSCs(n=3)500μl were injected via the caudal vein.And the myocardial infarction untreated group(n=3) and normal mice(n=3) fed with DOX-containing liquid at the same time were selected as control group 1 and control group 2.The heart function improvement was detected through cardiac color Doppler ultrasonography 72 h after providing the intervention measures.Normal mice with normal feeding(n=3) were selected as control group 3.The fluorescence intensity of heart region was detected by in vivo imaging of small animals.The local myocardial blood vessels in myocardial infarction were evaluated by tomato lectin,and the number of local C-kit positive cells in myocardial infarction was detected using immunohistochemistry.Results The expression rate of BMSCs CD29 in mice was 98.0％,the expression rate of CD44 was 100.0％,the expression rate of CD11b was 0.1％ and the expression rate of SCA-1 was 99.5％.The expression level of GATA-4 mRNA in GATA-4-BMSCs group was(78.17±1.32),which was higher than that in the negative control group (0.57±0.34) (t=2.576,P<0.05).The ejection fraction(EF) and central systolic(FS) of each group were compared,and the differences were statistically significant(P<0.05).The EF and FS changes in over-expressed GATA-4-BMSCs group were higher than those in the control group 1,control group 2,BMSCs group and empty vector-BMSCs group(P<0.05).The fluorescence intensity,the number of myocardial vessels and the number of C-kit positive cells in the heart region of each group were compared,and the differences were statistically significant(P<0.05).Among them,the fluorescence intensity in cardiac region,the number of C-kit positive cells in the over-expressed GATA-4-BMSCs group were higher than those in the control group 1,the control group 2,the control group 3,the BMSCs group and the empty vector-BMSCs group(P<0.05),and the number of myocardial vessels in the over-expressed GATA-4-BMSCs group were higher than those in the BMSCs group and the empty vector-BMSCs group(P<0.05).Conclusion Over-expressed GATA-4 BMSCs can promote the local vascular proliferation in the myocardial infarction and effectively mobilize the C-kit positive cells to repair myocardial injury through enhancing the"homing" effect.关键词
心肌梗死/GATA4转录因子/骨髓间充质干细胞Key words
Myocardial infarction/GATA4 transcription factor/Bone mesenchymal stem cells分类
医药卫生引用本文复制引用
贺继刚,韩金秀,李贝贝,李宏远,李洪荣,严丹..过表达GATA-4小鼠骨髓间充质干细胞修复心肌损伤的效果研究[J].中国全科医学,2018,21(2):167-172,6.基金项目
国家自然科学基金资助项目(81460073) （81460073）
云南省科技厅—昆明医科大学应用基础研究联合专项(2014FB089) （2014FB089）
云南省教育厅科学研究基金(2015Z051) （2015Z051）
中国博士后科学基金(2015M582764XB) （2015M582764XB）
成都医学院2015年度科研项目(CYZ15-18) （CYZ15-18）
