| 张鹭鹭刘惊今王永顺刘芳于波△.microRNA499 慢病毒载体转染诱导大鼠骨髓间充质干细胞
向心肌样细胞分化*[J].,2012,12(26):5027-2031 |
| microRNA499 慢病毒载体转染诱导大鼠骨髓间充质干细胞
向心肌样细胞分化* |
| miR-499 Induces Cardiac Differentiation of Rat Bone Marrow-DerivedMesenchymal Stem Cells* |
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| DOI： |
| 中文关键词: microRNA 499 骨髓间充质干细胞 心肌样分化 慢病毒载体转染 |
| 英文关键词: miR-499 Mesenchymal stem cells Cardiac differentiation Lentiviral vectors transfection |
| 基金项目:省部共建教育部重点实验室（哈尔滨医科大学）开放基金项目（KF201004) |
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| 中文摘要: |
| 目的：探讨microRNA 499( miR-499)慢病毒转染对诱导大鼠骨髓来源间充质干细胞(BM-MSCs)向心肌样细胞分化的作用。
方法：取第四代Wistar大鼠骨髓来源间充质干细胞进行流式细胞检测，鉴定干细胞表面特异标记物。使用符合干细胞鉴定标准的
细胞批次用于后续实验。实验设置miR499慢病毒转染、慢病毒空白转染2个处理组，分别于处理后即日、1d，3d，5d，7d 收集细胞
进行下列实验：实时荧光定量PCR 检测心肌重要转录因子GATA4、NKx2.5 和MEF2C 的mRNA 表达, western-blot 检测心肌特
异蛋白I( cTnI)的表达。结果：培养第四代Wistar大鼠骨髓来源间充质干细胞表达干细胞表面特异标记物，可用于实验。大鼠骨髓
来源间充质干细胞microRNA 499慢病毒载体转染后microRNA 499 表达明显升高，且转染后1d，3d，5d，7d, GATA4、NKx2.5 和
MEF2C的mRNA表达逐渐增强。慢病毒空白转染组未见明显变化。western-blot检测自第3天开始可见cTnI阳性表达条带，慢病
毒空白转染组未检测到明显阳性表达条带。结论：microRNA 499可诱导大鼠骨髓来源间充质干细胞向心肌样细胞分化。 |
| 英文摘要: |
| Objective: To investigate the effect of miR-499 on differentiation of cardiomyocyte-like cells from rat bone
marrow-derived mesenchymal stem cells (BM-MSCs). Methods: The fourth passage of rat BM-MSCs were infected with lentiviral vectors
bearing miR-499 or empty lentiviral vectors as control. The expression of cardiac-specific markers, NKx2.5, GATA4, MEF2C, and
cTnI in these cells were examined by real-time PCR and western blot. Results: The fourth passage of the cultured rat BM-MSCs express
significant markers of MSCs. After transfection of microRNA 499 in rat BM-MSCs, the expression level of microRNA 499 was markedly
enhanced. The mRNA expression of GATA4, NKx2.5 and MEF2C gradually increased from day 1 to day 7, the cTnI expression was
detected from day 3. While there were no significant changes of GATA4, NKx2.5 and MEF2C mRNA levels in BM-MSCs infected with
empty lentiviral vectors, and cTnI expression cannot be detected either. Conclusion: Over-expression of miR-499 rat BM-MSCs can
induce rat BM-MSCs toward cardiac differentiation. |
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