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目的 探讨破骨样细胞(OLC)及其亚细胞成分对成骨细胞(OB)Cbfα1表达的影响。方法C57雌性小鼠,经尾静脉注射5 -FU后,取其脾脏细胞,在白介素(IL) 3、IL 6和粒细胞巨噬细胞集落刺激因子(GM CSF)、1α, 25 (OH)2D3的诱导下获得大量OLC。OLC在骨片上培养即获得骨片上的OLC。NaF、两种OLC、离心去除OLC的培养基及其亚细胞结构—细胞核、线粒体与OB共培养5d后,分别使用半定量RT PCR和免疫组织化学的方法检测OBCbfα1mRNA和蛋白质的表达活性。结果 NaF、两种OLC的细胞核、线粒体、细胞浆和离心去除OLC后的培养基均可使OBCbfα1mRNA的表达明显加强(均P<0. 05);NaF,两种OLC的细胞浆和离心去除OLC后的50%培养基均可显著增加OBCbfα1蛋白质水平的表达(均P<0. 05)。结论 OLC的亚细胞成分和离心去除OLC后的培养基均对OBCbfα1表达具有促进作用,同时Cbfα1的表达存在多水平的调节机制。
Objective To investigate the effect of osteoclast-like cells (OLCs) and their subcellular components on the expression of Cbfα1 in osteoblasts (OB). Methods C57 female mice were injected with 5-FU via the tail vein and spleen cells were harvested. The spleen cells were collected and cultured in the presence of interleukin (IL) 3, IL 6 and GM CSF, 1α, 25 (OH) 2D3 A large number of OLCs were obtained under induction. OLC is obtained on the bone chip to obtain OLC on the chip. NaF, two kinds of OLC, and centrifugation to remove OLC, and its subcellular structure-nucleus, mitochondria and OB were cultured for 5 days. The expression of OBCbfα1 mRNA and protein were detected by semi-quantitative RT-PCR and immunohistochemistry respectively. Results The expression of OBCbfα1 mRNA in NaF, the nucleus, mitochondria, cytoplasm and cytosol in both OLC and OLC after centrifugation were significantly enhanced (all P <0.05); NaF, cytoplasm and centrifugal removal of OLC After 50% of the medium can significantly increase the expression of OBCbfα1 protein levels (P <0.05). Conclusion The subcellular components of OLC and the medium after centrifugation to remove OLC all promote the expression of OBCbfα1, and there is a multi-level regulation mechanism of Cbfα1 expression.