microRNAs (miRNAs) have recently been recognized as playing an important role in bone‐associated diseases. This study investigated whether the reduced miR‐155‐5p in steroid‐associated osteonecrosis of the femoral head (ONFH) attenuated osteogenic differentiation and cell proliferation by targeting GSK3B. Bone marrow was collected from the proximal femurs of patients with steroid‐associated ONFH (n = 10) and patients with new femoral neck fracture (n = 10) and mesenchymal stem cells (MSCs) were isolated. The expression profile, the biological function of miR‐155‐5p, and the interaction between miR‐155‐5p and GSK3B were investigated by cell viability measurement, western blot, real‐time polymerase chain reaction, luciferase reporter assay, and Alizarin Red S (ARS) staining of MSCs. The MSCs that were obtained from the femoral neck fracture group and from the steroid‐associated ONFH group were transfected with or without miR‐155‐5p. We found that, in ONFH samples, the level of mature miR‐155‐5p was significantly lower than that of control samples. By inhibiting GSK3B, miR‐155‐5p promoted the nuclear translocation of β‐catenin, increased the expression of osteogenesis‐related genes, and facilitated the proliferation and differentiation of MSCs. Restoring the expression of GSK3B in MSCs partially reversed the effect of miR‐155‐5p. These findings suggest that reduced miR‐155‐5p in steroid‐associated ONFH attenuates osteogenic differentiation and cell proliferation by increased levels of GSK3B and inhibition of Wnt signaling.

中文翻译：

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miR-155-5p通过靶向类固醇相关骨坏死中的GSK3B来调节间充质干细胞成骨和增殖

microRNA（miRNA）最近被认为在骨相关疾病中起重要作用。本研究调查了类固醇相关股骨头坏死 (ONFH) 中 miR-155-5p 的减少是否通过靶向 GSK3B 减弱了成骨分化和细胞增殖。骨髓是从患者的近侧股骨收集与类固醇相关的股骨头坏死（Ñ  = 10）和患有新股骨颈骨折（Ñ = 10) 和间充质干细胞 (MSC) 被分离出来。通过细胞活力测量、蛋白质印迹、实时聚合酶链反应、荧光素酶报告基因检测和茜素红 S 研究了表达谱、miR-155-5p 的生物学功能以及 miR-155-5p 与 GSK3B 之间的相互作用(ARS) MSCs 染色。从股骨颈骨折组和类固醇相关 ONFH 组获得的 MSCs 转染有或没有 miR-155-5p。我们发现，在 ONFH 样本中，成熟 miR-155-5p 的水平显着低于对照样本。通过抑制 GSK3B，miR-155-5p 促进 β-catenin 的核转位，增加成骨相关基因的表达，促进 MSCs 的增殖和分化。恢复 GSK3B 在 MSC 中的表达部分逆转了 miR-155-5p 的作用。这些发现表明，类固醇相关 ONFH 中 miR-155-5p 的减少通过增加 GSK3B 水平和抑制 Wnt 信号来减弱成骨分化和细胞增殖。