目的 初步探讨骨桥蛋白对破骨细胞体外分化的影响。方法 体外培养小鼠破骨前体RAW264.7细胞,RANKL诱导刺激其分化为破骨细胞,并加入骨桥蛋白进行干预。MTT法及TRAP染色分别检测破骨细胞细胞增殖及前体破骨细胞分化情况,RT-PCR检测RANK的mRNA表达水平。结果 骨桥蛋白可明显促进RAW264.7细胞向破骨细胞分化、增加细胞增殖,与对照组相比差异具有统计学意义(P＜0.05),并使破骨细胞特征性基因RANK的mRNA表达上调约2.67倍。结论 骨桥蛋白可能通过影响RANK/RANKL信号通路促进RAW264.7细胞分化为破骨细胞,有望成为防治骨质疏松等骨相关疾病的新靶点。
Abstract
Objective To study the effect of osteopontin (OPN) on differentiation of osteoclasts (OC) by RANKL in vitro. Methods Mouse osteoclasts precursor cell line RAW264.7 was cultured and induced to form OC, and was added with osteopontin (OPN). Then, the proliferation and differentiation of OC were detected by MTT assay and TRAP staining. Furthermore, RT-PCR was used to measure the gene expression of RANK. Results The model of OC in vitro was established successfully, accompanied by the significant increase of proliferation and differentiation of OC (P<0.05). Meanwhile, OPN promoted the expression of RANK mRNA in OC obviously, which could be increased 2.67 times. Conclusions OPN affects the differentiation of RAW264.7 cells to form OC via RANK/ RANKL pathway, which may provide a new target for the prevention and treatment of osteoporosis and bone-related diseases.
关键词
骨桥蛋白 /
RAW264.7细胞 /
破骨细胞 /
细胞分化
Key words
osteopontin /
RAW264.7 cells /
osteoclasts /
differentiation
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参考文献
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基金
国家自然科学基金(81202843),天津市应用基础与前沿技术研究计划(14JCZDJC36700),武警后勤学院科研基金项目(WHJ201405)
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