论文部分内容阅读
目的观察不同浓度NaF(0mg/L、2 mg/L、10 mg/L、20 mg/L、40 mg/L、60 mg/L、80 mg/L)在不同时间(24 h、48 h、72 h),对RAW264.7细胞活力及破骨活性的影响。方法 RAW264.7细胞浓度为5×104个/ml接种24孔板,每孔加2 ml培养基;按NaF浓度和时间分组,作MTT,用酶标仪测吸光度值,判断细胞活力;免疫组织化学法测定破骨细胞内基质金属蛋白酶9(MMP-9)和组织蛋白酶K的表达。结果 NaF浓度变量和时间变量交互作用对细胞活力(吸光度值)造成显著的影响,细胞活力随NaF浓度升高依次呈梯度样显著降低;NaF浓度为20 mg/L时,RAW264.7细胞活力未见明显抑制;随着NaF作用时间延长,RAW264.7细胞活力依次呈显著升高趋势;MMP-9表达随氟剂量增高而增强,组织蛋白酶K表达各组未见差异。结论 NaF对RAW264.7细胞活力的抑制是暂时性的,随着时间的延长NaF对RAW264.7细胞活力的抑制作用减弱;氟通过提高破骨细胞内MMP-9的表达来增强破骨细胞骨吸收活性。
Objective To observe the effects of NaF at different concentrations (0, 2, 10, 20, 40, 60 mg / L, 80 mg / L) 72 h) on the viability and osteoclast activity of RAW264.7 cells. Methods RAW264.7 cells were inoculated into 24-well plates at a concentration of 5 × 104 cells / ml and supplemented with 2 ml of medium per well. MTT was performed by concentration and time of NaF. The absorbance was measured by microplate reader to determine cell viability. The chemical method was used to determine the expression of matrix metalloproteinase 9 (MMP-9) and cathepsin K in osteoclasts. Results The interaction of NaF concentration variables and time variables had a significant effect on the cell viability (absorbance value). The viability of cells decreased markedly with the increase of NaF concentration. When the concentration of NaF was 20 mg / L, the viability of RAW264.7 cells See obvious inhibition; With the prolongation of NaF time, vitality of RAW264.7 cells followed by a significant upward trend; MMP-9 expression increased with increasing fluoride dose, no difference in the expression of cathepsin K in each group. CONCLUSIONS: The inhibitory effect of NaF on the viability of RAW264.7 cells is transient. The inhibitory effect of NaF on the viability of RAW264.7 cells is weakened with time. Fluorine enhances the expression of MMP-9 in osteoclasts to enhance osteoclast Absorb activity.