MiR-291a-3p regulates the BMSCs differentiation via targeting DKK1 in dexamethasone-induced osteoporosis
MiR-291a-3p regulates the BMSCs differentiation via targeting DKK1 in dexamethasone-induced osteoporosis
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DOI：
10.1002/kjm2.12134
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发表时间：
2020-01-01
影响因子：
3.3
通讯作者：
Wu, Yu-Chi
中科院分区：
文献类型：
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作者：
Li, Zhe-Hai;Hu, He;Wu, Yu-Chi
骨质疏松症是一种骨骼疾病，影响55%的60岁以上的人，由于人口老龄化，这一数字仍在增加。预防骨质疏松症的一种方法是增加新骨的形成，同时防止旧骨的吸收。因此，骨髓间充质干细胞（BMSCs）的成骨分化对于改善骨质疏松症的治疗具有重要意义。另一方面，糖皮质激素（GCs）被广泛用于治疗慢性炎症性疾病，但长期暴露于GCs可诱发骨质疏松症。在本研究中，我们用地塞米松（DEX）处理BMSCs来模拟GC诱导的骨质疏松症。采用MTT法、ALP活性测定和茜素红染色检测miRNA-291 a-3 p在DEX诱导的成骨分化抑制中的作用。进一步采用qPCR和western blot方法研究miRNA-291 a-3 p对BMSCs分化的影响。结果表明，miRNA-291 a-3 p能提高DEX抑制的BMSCs的细胞活力、成骨分化和ALP活性。此外，我们发现当转染miRNA-291 a-3 p模拟物时，成骨基因Runx 2、DMP 1和ALP上调，而脂肪生成基因C/EBP α和PPAR γ下调。此外，我们还证实了miRNA-291 a-3 p通过直接抑制DKK 1 mRNA和蛋白的表达，进而激活Wnt/beta-catenin信号通路，促进BMSCs的成骨分化。我们的研究表明，miR-291 a-3 p在预防骨质疏松症中起着重要作用，并可能作为一个潜在的miRNA骨质疏松症生物标志物。
Osteoporosis is a skeleton disease affecting 55% of people over age 60, and the number is still increasing due to an ageing population. One method to prevent osteoporosis is to increase the formation of new bone while preventing the resorption of older bone. Thus, osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) is of great importance in improving the treatment of osteoporosis. On the other hand, glucocorticoids (GCs) are widely used to treat the chronic inflammatory disorders, but long-term exposure to GCs can induce osteoporosis. In present study, we treated BMSCs with dexamethasone (DEX) to simulate GC-induced osteoporosis. MTT assay, ALP activity, and Alizarin Red were used to evaluate the role miRNA-291a-3p in the DEX-induced osteogenic differentiation suppression. Further, we used qPCR and western blot to investigate the mechanisms of miRNA-291a-3p affecting BMSCs differentiation. The results showed that miRNA-291a-3p could improve the cell viability, osteogenic differentiation, and ALP activity, which are suppressed by DEX in BMSCs. Furthermore, we found that the osteogenesis genes Runx2, DMP1, and ALP were upregulated whereas the lipogenic genes C/EBP alpha and PPAR gamma were downregulated when miRNA-291a-3p mimics were transfected. Additionally, we demonstrated that miRNA-291a-3p promoted BMSCs' osteogenic differentiation by directly suppressing DKK1 mRNA and protein expression and subsequently activating Wnt/beta-catenin signaling pathway. Our study suggests that miR-291a-3p plays an important role in preventing osteoporosis and may serve as a potential miRNA osteoporosis biomarker.