摘要
目的检测微小RNA（miRNA，miR）-27a调控过氧化物酶体增殖子活化受体-γ（PPAR-γ）和骨形态发生蛋白-2（BMP-2）对骨髓间充质干细胞（BMSCs）分化的影响。方法将培养的40只大鼠BMSCs，随机分4组，正常对照组：细胞不作特殊处理；模型组：细胞给予1×10-7mol／L地塞米松；无关序列组：将无关序列基因电转入细胞，给予1×10 -7mol／L地塞米松；实验组：将具有双向靶向作用的miR-27a电转入细胞，给予1×10-7mol／L地塞米松。采用实时荧光定量聚合酶链反应（RT—qPCR）技术测定PPAR-γ和BMP-2mRNA的相对表达量。结果处理细胞7d时，实验组细胞中PPAR-γmRNA的相对表达量（1．203±0．111）较模型组（1．877±0．225）、无关序列组（1．913±0．195）明显降低（P〈0．05），近似于正常组（1．000）且与其差异无统计学意义（P〉0．05）。实验组细胞中BMP-2mRNA的相对表达量（0．832±0．105）较模型组（0．455±0．051）、无关序列组（0．422±0．038）明显升高（P〈0．05），近似于正常组（1．000）且与其差异无统计学意义（P〉0．05）。结论miR-27a能够有效抑制PPAR-γ表达，维持BMP-2表达。
Objective To explore the effect on the differentiation of steroid- induced bone marrow mesenchymal stem cells （BMSCs） of rats by tarteting regulation of microRNA （miRNA, miR） -27a on per- oxisome proliferator - activated receptor γ, （ PPAR -γ） and bone morphogenetic protein - 2 （ BMP - 2 ）. Methods BMSCs of 40 rats were expanded and randomly divided into 4 groups. In normal control group, the ceils were not treated. In model group, the cells were treated with 1 0133× 10 -7 mol/L dexamethasone. In irrelative sequence group, the cells were electroporated with the irrelative sequence that was ineffective at targeting the PPAR -7 gene, and treated with 1 x 10-7 mol/L dexamethasone. In experimental group, the ceils were electroporated with miR - 27a and treated with 1 ~ 10-7 mol/L dexamethasone. The real - time quantitative reverse transcription polymerase chain reaction （RT -qPCR） was used to detected the expression of PPAR - ~/ and BMP - 2 mRNA R^sults At 7th day after treatment, the expression level of PPAR - ~/ mRNA in experimental group （ 1. 203 ± 0. 111 ） was significantly lower than that in model group （ 1. 877 -± 0. 225） and irrelative sequence group （1. 913 ± 0. 195 ） （P 〈 0.05 ）, and was similar to that in normal control group （ P 〉 0.05 ）. The expression level of BMP - 2 mRNA in experimental group （0. 832 ± 0. 105 ） was significantly higher than that in model group （0. 455 ±± 0. 051 ） and irrelative sequence group （ 0. 422 ± 0. 038 ） （ P 〈 0. 05 ）, and was similar to that in normal control group （ P 〉 0. 05 ）. Conclusion The miR- 27a may inhibit the expression of PPAR -%, effectively and maintain the expression of BMP- 2.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2015年第12期3110-3112,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（81470106）
