微小RNA-548d-5p靶向调控过氧化物酶体增殖物激活受体γ对乙醇诱导大鼠骨髓间充质干细胞分化的影响

Effect on differentiation of alcohol - induced bone marrow mesenchymal stem cells in rats with os- teonecrosis of femoral head by targeting regulation of microRNA - 548d - 5p on expression of peroxisome proliferator activated receptor γ

目的观察微小RNA（miRNA，miR）-548d-5p靶向调控过氧化物酶体增殖物激活受体吖（PPARl）基因表达对乙醇诱导大鼠骨髓间充质干细胞（BMSCs）分化的影响。方法2个月龄雄性大鼠制备BMSCs。将细胞实验分4组：（1）正常对照组：细胞不做特殊处理；（2）模型组：给予细胞0．09mol／L乙醇；（3）无关序列组：将无关序列电转入细胞，给予细胞0．09mol／L乙醇；（4）基因组：将miR-548d-5p电转入细胞，给予0．09mol／L乙醇。在细胞被乙醇诱导第3天和第7天，采用TaqMan反转录-聚合酶链反应（1iT—PCR）方法检测各组细胞PPARγmRNA。结果乙醇诱导第3天，正常对照组、模型组、无关序列组、基因组中PPARγmRNA表达量分别为1．000、1．720±0．176、1．757±0．188、1．017±0．096。第7天，4组中PPAR-γmRNA表达量分别为1．000、1．806±0．209、1．829±0．223、1．118±0．108，基因组中PPARγ mRNA表达低于模型组和无关序列组（P〈0．05），接近于正常组且与其差异无统计学意义（P〉0．05）。模型组和无关序列组中PPARγ mRNA表达高于正常对照组（P〈0．05）。结论miR-548d-5p能够抑制乙醇诱导BMSCs内PPARγ mRNA的表达。

Objective To explore the effect on the differentiation of alcohol -induced bone marrow mesenchymal stem cells （BMSCs） in rats by targeting regulation of microRNA （miRNA, miR） -548d -5p on peroxisome proliferator activated receptor γ （PPARγ）. Methods BMSCs were obtained from 2 - month - old male rats. The experiment was divided into 4 groups. In normal control group, the cells were not trea- ted. In model group, the cells were treated with 0.09 mol/L alcohol. In irrelative sequence group, the cells were electroporated with the irrelative sequence that was ineffective at targeting the PPART, and trea- ted with O. 09 mol/L alcohol. In gene group, the cells were electroporated with miR -548d -5p and trea- ted with O. 09 mol/L alcohol. The expression of PPARγ mRNA was determined using the method of Taq- Man reverse transcriptase - polymerase chain reaction（ RT - PCR） at 3rd and 7th day. Results At 3rd day after treatment, the expression level of PPARγ mRNA in normal control group, model group, irrelative se- quence group and gene group was 1. 000, 1. 720 ±0. 176, 1. 757 ±0. 188, and 1. 017 ±0. 096 respectively. At 7th day, the expression level of PPARγ mRNA in in normal control group, model group, irrelative sequence group and gene group was 1. 000, 1. 806 ±0. 209, 1. 829 0. 223, and 1. 118 ±0. 108 respectively. The expression level of PPARγmRNA in gene group was lower than that in model group and irrela- tire sequence group （ P 〈 0. 05 ）, and similar to that in normal control group （ P 〉 0. 05 ）. The expression level of PPARγ mRNA was higher in model group and irrelative sequence group than that in normal control group, P 〈 0. 05. Conclusion miR - 548d - 5p can inhibit the expression of PPARγmRNA.