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目的:探讨不同浓度的无机磷液对体外培养MC3T3-E1细胞的成骨和破骨相关基因mRNA表达的影响。方法:体外常规培养MC3T3-E1细胞。根据所用培养液外加无机磷浓度的不同,将实验细胞分为4组:0 mM组(对照组)、1 mM组、3 mM组和5 mM组。每组设3个复孔,培养48 h后收取细胞标本,通过Real-time PCR检测MC3T3-E1细胞成骨和破骨相关基因mRNA的表达。结果:与对照组相比,成骨相关基因ALP、BSP、OC及OSXmRNA的表达在1 mM组和3 mM组增加,而在5 mM组则减少,差异具有统计学意义(P<0.05);成骨抑制基因SOST及破骨相关基因CTSK、TRAPmRNA的表达,在1 mM组、3 mM组和5 mM组都增加,差异具有统计学意义(P<0.05)。结论:高浓度(5 mM)的无机磷环境对MC3T3-E1细胞的成骨相关基因表达起显著抑制作用,对破骨相关基因表达起显著增强作用,提示高浓度无机磷抑制MC3T3-E1细胞的成骨活性。
OBJECTIVE: To investigate the effect of different concentrations of inorganic phosphate on osteogenic and osteoclastic related gene mRNA expression in MC3T3-E1 cells cultured in vitro. Methods: MC3T3-E1 cells were cultured in vitro routinely. The experimental cells were divided into 4 groups according to the culture medium plus inorganic phosphorus concentration: 0 mM group (control group), 1 mM group, 3 mM group and 5 mM group. Three replicate wells were established in each group. Cell samples were collected 48 h after culture. The mRNA expression of osteoclasts and osteocarst related genes in MC3T3-E1 cells was detected by Real-time PCR. Results: Compared with the control group, the expressions of ALP, BSP, OC and OSX mRNA increased in 1 mM and 3 mM groups and decreased in 5 mM group, with statistical significance (P <0.05). The expression of osteosarcoma suppressor gene SOST and osteoclastic related gene CTSK and TRAP mRNA increased in 1 mM, 3 mM and 5 mM groups, with statistical significance (P <0.05). CONCLUSION: High concentration (5 mM) inorganic phosphorus can significantly inhibit osteoblast-related gene expression in MC3T3-E1 cells and significantly enhance osteoclast-related gene expression, suggesting that high concentrations of inorganic phosphorus inhibit MC3T3-E1 cells Osteogenic activity.