促红细胞生成素在成骨细胞分化中的实验研究

Roles of erythropoietin on the differentiation of osteoblasts

目的研究促红细胞生成索(EPO)对成骨细胞分化和功能的影响以及EPO-EPO受体(EPOR)价号在成骨细胞中的作用,探讨EPO在骨稳态中的作用及其机制。方法体外培养小鼠骨髓基质细胞系(ST2),加入EPO蛋白,采用实时荧光定量聚合酶链式反应方法检测成骨分化相关基因的变化,采用碱性磷酸酶(ALP)和茜素红染色方法观察成骨细胞骨形成功能。采用RNA干扰技术沉默EPOR基因,检测阻断EPO-EPOR信号通路后成骨细胞分化和功能的变化。结果骨髓基质细胞表达EPOR,EPO与受体EPOR结合后,可诱导其向成骨细胞分化,增加Runx2、Alp和Col1等成骨细胞相关基因的表达,同时增加ALP蛋白的表达和钙结节的沉积。EPOR基因沉默后,成骨相关基因的表达受到抑制。结论 EPO通过EPOR信号促进成骨细胞的分化和功能。

Objective This study aimed to investigate the effects of erythropoietin（EPO） on the differentiation and function of osteoblasts, to identify the roles of EPO-erythropoietin receptor（EPOR） in osteoblasts, and to explore the effects and mechanism of EPO in bone homeostasis. Methods Mouse bone marrow stromal cell line ST2 was cultured in vitro and treated with EPO. Changes in osteoblastic gene levels were quantified by real-time polymerase chain reaction. The function of osteoblasts in bone formation was observed by alkaline phosphatase（ALP） and alizarin red staining. EPOR expression was knocked down by RNA interfere. The differentiation and function of osteoblasts were then tested by the above methods. Results ST2 ceils expressed EPOR. After binding with EPOR, EPO induced osteoblast differentiation from bone marrow stromal cells. Expression levels of osteoblast-related genes Runx2, Alp, and Coll increased. ALP and calcium deposition increased. Expression levels of osteoblast-related genes were repressed after EPOR knockdown. Conclusion EPO promotes the differentiation and function of osteoblasts through EPO-EPOR signaling.