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目的探讨miR-29b在骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)分化为软骨细胞时,对细胞肥大化进程的调控作用。方法培养BMSCs并进行软骨细胞肥大化方向的诱导,检测软骨细胞标志基因Ⅱ型胶原(ColⅡ)、肥大化细胞标志基因Ⅹ型胶原(ColⅩ和Runx2),并用特异性染色法确定细胞肥大化诱导成功,再检测miR-29b在转录水平的表达情况。然后将实验分为miR-29b过表达(miR-29b mimics)和miR-29b抑制(miR-29b抑制剂)两组,每组分别设置空白对照组,分别检测两处理组中上述软骨细胞和肥大化细胞相关标志的表达变化。结果 miR-29b过表达组中,ColⅡ表达与对照组相比显著降低(P<0.05),ColⅩ和Runx2表达显著升高(P<0.05);而miR-29b抑制组中,ColⅡ的表达与对照组相比显著升高(P<0.05),ColⅩ和Runx2显著降低(P<0.05)。结论miR-29可以有效促进BMSCs向肥大化的软骨细胞分化,而抑制miR-29b可降低BMSCs来源的软骨细胞的肥大化。
Objective To investigate the regulatory effect of miR-29b on the process of cell hypertrophy when differentiated into chondrocytes by bone marrow mesenchymal stem cells (BMSCs). Methods BMSCs were cultured and induced by chondrocyte hypertrophy. The expression of collagen Ⅱ (Col Ⅱ) and mast cell marker gene Ⅹ (Colx) and chondrocyte Runx2 were detected by specific staining and the successful induction of cell hypertrophy , Then detect the expression of miR-29b at the transcriptional level. Then the experiment was divided into two groups: miR-29b mimics and miR-29b inhibitor (miR-29b inhibitor), each group was set up blank control group, respectively, to detect the two groups of chondrocytes and hypertrophy Changes in the expression of chemokines. Results In the miR-29b overexpression group, the expression of ColⅡ was significantly lower than that of the control group (P <0.05), and the expression of Col X and Runx2 was significantly increased (P <0.05); while in the miR-29b inhibition group, (P <0.05), while Col X and Runx2 decreased significantly (P <0.05). Conclusion miR-29 can effectively promote the differentiation of BMSCs into hypertrophic chondrocytes, while inhibiting miR-29b can reduce the proliferation of BMSCs-derived chondrocytes.