补肾方含药血清处理的大鼠成骨细胞miRNA差异表达

Differentially Expressed miRNAs in Rat Osteoblasts Treated by Serum Containing Bushen Prescription

目的筛选补肾方含药血清干预的大鼠成骨细胞中差异表达的miRNA,为补肾方治疗骨质疏松症提供可供选择的目标miRNA,以探讨其通过调控靶基因表达发挥治疗效应的机制。方法经细胞性状鉴定的原代大鼠成骨细胞用含药血清干预,采用miRNA芯片检测补肾方含药血清干预3d与干预1d的成骨细胞miRNA表达水平,并挑选差异表达2倍以上的miRNA进行定量PCR验证。结果培养的原代细胞具有典型的成骨细胞性状特征。与对照血清相比,补肾方含药血清干预3d的成骨细胞共有24个2倍以上表达差异的miRNA,其中上调的16个,下调的8个。差异表达2倍以上的miRNA定量PCR结果与芯片数据一致。结论补肾方通过成骨细胞中差异表达的miRNA调控靶基因改善骨质疏松症。

Objective To investigate the differentially expressed miRNAs in rat osteoblasts treated by serum containing Bushen Prescription,the target miRNAs for the treatment of osteoporosis with Bushen Prescription, and mechanisms for the therapeutic effect of Bushen Prescription by regulating the expression of target genes.Methods Primary rat osteoblasts were treated by serum containing Bushen Prescription after identification.The miRNA microarray was used to measure the expression of miRNAs in osteoblasts treated by serum containing Bushen Prescription for 1 or 3 days, and real-time quantitative PCR was used to further validate the miRNAs which were 〉2-fold differentially expressed between the two groups.Results The primary culturedosteoblasts had the typical properties of osteoblasts.A total of 24 miRNAs were 〉2-fold differentially expressed between the two groups, and compared with the control serum group, the serum containing Bushen Prescription had 16 up-regulated miRNAs and 8 down-regulated miRNAs.The real-time PCR results for these 24 miRNAs were consistent with the microarray results.Conclusion Bushen Prescription can regulate target genes via differentially expressed miRNAs in osteoblasts and thus improve osteoporosis.