CAJ | 학술논문

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OyCl3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1×10-5 mol·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1×10-5 mol·L-1的DyCl3促进成骨细胞矿化结节的形成,而浓度为1×10-7 mol·L-1和1×10-6 mol·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1×10-8 mol·L-1它则对成骨细胞矿化功能没有影响.浓度1×10-6 mol-L-1的DyCl3显著降低PPAR-γ mRNA表达水平,但相同浓度DyCl3则显著上调RUNX-2 mRNA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.
채용MTT법、감성린산매활성측정、유홍O적염색화정량측정、광화공능적측정이급qRT-PCR등수단재세포화분자수평상연구료OyCl3대원대배양적성골세포증식、분화화광화공능적영향.연구결과표명,재측시농도범위내,DyCl3균억제성골세포증식.농도위1×10-8,1×10-6화1×10-5 mol·L-1적DyCl3촉진성골세포분화.재측시농도범위내,DyCl3균억제성골세포횡향분화위지방세포.농도위1×10-5 mol·L-1적DyCl3촉진성골세포광화결절적형성,이농도위1×10-7 mol·L-1화1×10-6 mol·L-1적DyCl3억제성골세포광화결절적형성,진일보강저농도위1×10-8 mol·L-1타칙대성골세포광화공능몰유영향.농도1×10-6 mol-L-1적DyCl3현저강저PPAR-γ mRNA표체수평,단상동농도DyCl3칙현저상조RUNX-2 mRNA표체수평.실험결과제시,DyCl3대체외배양적성골세포증식、분화급광화공능적영향여농도화작용시간유관,이차,타문시영향기생물효응종독성도활성,종손상도보호,종상조도하조전변적관건인소.저사결과대심입리해희토리자대골대사적영향구유중요적개치.
A series of experimental methods including 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide(MTT)test,alkaline phosphatase(ALP)activity measurement,Oil Red O stain and measurement,mineralized function expression and quantitive real time RT-PCR(qRT-PCR)were employed to assess the effects of DyCl3 on the the proliferation,differentiation and mineralization function of primary osteoblasts(OBs)in vitro at cell and molecular levels.The results showed that DyC13 inhibited the proliferation of OBs at tested concentrations.As to the differentiation of OBs,DyCl3 promoted the differentiation of OBs at concentrations of 1×10-8,1×10-6,and 1×10-5 mol·L-1.DyCl3 suppressed the adipogenic transdifferentiation of OBs at tested concentrations.DyCl3 promoted the formation of mineralized matrix nodules of OBs at a higher concentration 1×10-5 mol·L-1,turned to inhibit the formation of mineralized matrix nodules of O Bs at concentrations of 1 × 10-7 and 1 × 10-6 mol·L-1,but had no effect on the formation of mineralized matrix nodules of OBs at a concentration of 1x10-8 mol.L-1.The expression of the mRNA for PPAR-γ was significantly down-regulated,the expression of the mRNA for RUNX-2 was up-regulated in the presence of 1 ×10-6 mol·L-1 DyCl3.Our experimental results suggest that the effects of DyCl3 on the proliferation,differentiation and mineralization function of OBs in vitro depend on the concentration and culture time,moreover, they are pivotal factors for switching the biological effects of rare earth ions from toxicity to activity,from damage to protection,or from down-regulation to up-regulation.These results are very helpful for elucidating deeply the effects of rare earth ions on bone metabolism.