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采用MTT法、碱性磷酸酶活性测定、油红O的染色和定量测定、矿化功能的测定以及qRT-PCR等手段在细胞和分子水平上研究了DyCl3对原代培养的成骨细胞增殖、分化和矿化功能的影响。研究结果表明,在测试浓度范围内,DyCl3均抑制成骨细胞增殖。浓度为1×10-8,1×10-6和1×10-5mol·L-1的DyCl3促进成骨细胞分化。在测试浓度范围内,DyCl3均抑制成骨细胞横向分化为脂肪细胞。浓度为1×10-5mol·L-1的DyCl3促进成骨细胞矿化结节的形成,而浓度为1×10-7mol·L-1和1×10-6mol·L-1的DyCl3抑制成骨细胞矿化结节的形成,进一步降低浓度为1×10-8mol·L-1,它则对成骨细胞矿化功能没有影响。浓度1×10-6mol·L-1的DyCl3显著降低PPAR-γmRNA表达水平,但相同浓度DyCl3则显著上调RUNX-2mRNA表达水平。实验结果提示,DyCl3对体外培养的成骨细胞增殖、分化及矿化功能的影响与浓度和作用时间有关,而且,它们是影响其生物效应从毒性到活性,从损伤到保护,从上调到下调转变的关键因素。这些结果对深入理解稀土离子对骨代谢的影响具有重要的价值。
The effects of DyCl3 on the proliferation of primary cultured osteoblasts were investigated by MTT assay, alkaline phosphatase activity assay, staining and quantitation of oil red O, determination of mineralization function and qRT-PCR. Effects of differentiation and mineralization. The results indicate that DyCl3 inhibits osteoblast proliferation over the range of concentrations tested. DyCl3 at concentrations of 1 × 10 -8, 1 × 10 -6 and 1 × 10 -5 mol·L -1 promoted osteoblast differentiation. DyCl3 inhibited osteoblasts to differentiate into adipocytes laterally in the concentration range tested. DyCl3 at a concentration of 1 × 10-5mol·L-1 promoted the formation of mineralized nodules in osteoblasts, whereas DyCl3 at concentrations of 1 × 10-7mol·L-1 and 1 × 10-6mol·L-1 inhibited The formation of mineralized nodules in osteoblasts further reduced the concentration of 1 × 10-8 mol·L-1, which had no effect on the mineralization of osteoblasts. DyCl3 at a concentration of 1 × 10-6mol·L-1 significantly decreased the expression of PPAR-γmRNA, but the same concentration of DyCl3 up-regulated the expression of RUNX-2mRNA. The experimental results suggest that the effect of DyCl3 on the proliferation, differentiation and mineralization of cultured osteoblasts in vitro is related to concentration and duration of action, and that they affect their biological effects from toxicity to activity, from injury to protection, from up to down The key factor for change. These results are of great value for understanding the effects of rare earth ions on bone metabolism.