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目的探讨促甲状腺激素(TSH)对小鼠单核/巨噬细胞系RAW264.7细胞诱导分化为破骨样细胞过程的影响。方法体外培养小鼠单核/巨噬细胞系RAW264.7细胞,以破骨细胞分化因子(RANKL)诱导分化,同时加用不同浓度TSH处理7 d,抗酒石酸酸性磷酸酶(TRAP)染色鉴定并计数阳性细胞数量,采用Real-time PCR方法检测分化标志蛋白TRAP、基质金属蛋白酶-9(MMP-9)及组织蛋白酶K(Cathepsin K)的基因表达。结果RAW264.7细胞可在RANKL诱导下分化为破骨样细胞,高表达TRAP、M M P-9和Cathepsin K基因,TSH可剂量依赖性抑制细胞分化并下调上述基因的表达。结论 TSH可抑制破骨细胞的分化,对于维持骨量可能具有重要作用。
Objective To investigate the effect of thyroid stimulating hormone (TSH) on the differentiation of mouse monocyte / macrophage cell line RAW264.7 into osteoclast-like cells. Methods Mouse monocyte / macrophage cell line RAW264.7 cells were cultured in vitro and induced by osteoclast differentiation factor (RANKL). At the same time, different concentrations of TSH were added to treat RAW264.7 cells for 7 days. The cells were stained with tartrate - resistant acid phosphatase (TRAP) The number of positive cells was counted. Gene expression of TRAP, MMP-9 and Cathepsin K were detected by Real-time PCR. Results RAW264.7 cells could differentiate into osteoclast-like cells under the induction of RANKL, highly expressing TRAP, M M P-9 and Cathepsin K genes. TSH could inhibit cell differentiation and down-regulate the expression of these genes in a dose-dependent manner. Conclusion TSH can inhibit osteoclast differentiation and may play an important role in maintaining bone mass.