摘要
背景:最近发现,miR-378能调节骨髓间充质干细胞的抗缺氧能力,减轻其在缺氧环境下的凋亡。目的:观察miR-378过表达骨髓间充质干细胞在大鼠心肌梗死模型中的治疗作用。方法:体外培养至第3代的大鼠骨髓间充质干细胞用反转录病毒感染建立miR-378过表达细胞系。选用成年SD大鼠,结扎前降支制作急性心肌梗死模型,随机分为3组:对照组(n=10)、BMSC^(null)组(n=16)、BMSC^(miR-378)组(n=16)。对照组仅注射50μL生理盐水,后2组梗死局部注射50μL生理盐水中含1×10~7个空病毒转染和miR-378模拟物转染的骨髓间充质干细胞。移植24 h后,利用TUNEL染色测定移植骨髓间充质干细胞的凋亡率,Western blotting检测移植区域内血管内皮生长因子、转化生长因子β1蛋白表达。移植4周后,利用心脏超声评价心功能,取局部心肌组织行组织学及分子生物学分析。结果与结论:(1)移植24 h后,BMSC^(miR-378)组中凋亡细胞明显少于BMSC^(null)组(n=6,P<0.001),血管内皮生长因子、转化生长因子β1表达水平明显高于BMSC^(null)组(n=6,P<0.001);(2)移植4周后,BMSC^(miR-378)组中存活和分化为心肌细胞的骨髓间充质干细胞数量明显多于BMSC^(null)组(n=10,P<0.001)。与其他2组相比,BMSC^(miR-378)组中新生血管数量明显增多(P<0.001);心肌梗死面积明显减小(P<0.001);左室射血分数明显增加(P<0.05);左室舒张末容积明显缩小(P<0.05)。BMSCnull组中上述参数明显优于对照组(P<0.05);(3)结果表明,miR-378过表达可提高移植骨髓间充质干细胞的抗缺氧能力,继而促进心肌修复。
BACKGROUND： Recently, miR-378 has been shown to modulate the anti-hypoxia capacity of bone marrow mesenchymal stem cells（BMSCs） and reduce cell apoptosis under hypoxic conditions. OBJECTIVE： To investigate the benefits of miR-378-upregulated BMSC transplantation in a rat model of acute myocardial infarction.METHODS：Primary rat BMSCs were cultured in vitro.Until passage 3,the cells were infected with the lentivirus carrying synthetic miR-378 gene fragments.A rat model for acute myocardial infarction was constructed by ligating the left anterior descending artery.Thereafter,the animals were randomly assigned to three groups：control group（n=10）,BMSCnull group（n=16）and BMSCmiR-378 group（n=16）.In the latter two groups,50μL of normal saline containing 1×107 empty virus-transfected or miR-378-transfected BMSCs was injected into the region of myocardial infarction,respectively.Only 50μL of normal saline was injected in the control group.Twenty-four hours later,the apoptosis of transplanted BMSCs was evaluated with TUNEL,and expression level of vascular endothelial growth factor and transforming growth factor-βwas detected using western blot assay.Four weeks after treatment,the left ventricular function of rats was assessed by echocardiography,and then histological and molecular biology analyses were performed.RESULTS AND CONCLUSION：At 24 hours postoperatively,there were less apoptotic BMSCs and higher expression levels of vascular endothelial growth factor and transforming growth factor-βin the BMSCmiR-378 group than in the BMSCnull group（n=6,P〈0.001）.Four weeks later,there were more transplanted BMSCs and BMSCs-derived cardiomyocytes in the BMSCmiR-378 group than the BMSCnull group（n=10,P〈0.001）.Moreover,increased new vessel density（P〈0.001）,decreased infarcted area（P〈0.001）,preserved left ventricular ejection fraction（P〈0.05）,reduced left ventricular end-diastolic volume（P〈0.05）were found in the BMSCmiR-378 group,compared with the other two groups.The a
出处
《中国组织工程研究》
CAS
北大核心
2017年第9期1390-1396,共7页
Chinese Journal of Tissue Engineering Research
基金
辽宁省科学技术计划项目(2012225084)
沈阳市科学技术项目(F12-193-9-03
F16-206-9-08)
辽宁省教育厅科学研究一般项目(L2013316)~~
