【摘要】 目的观察不同浓度的3,3’,5-碘-L-甲腺原氨酸(3,3’,5-triiodo-L-thyronine,T3)对小鼠颅顶前成骨细胞亚克隆14 (MC3T3-E1 subclone 14)细胞增殖及分化水平的作用。方法分别给予不同浓度T3干预细胞,培养1、2、3 d后采用CCK8法检测细胞增殖水平;在成骨分化诱导试剂干预7 d基础上,结合上述T3给药,通过碱性磷酸酶(alkaline phosphatase,ALP)染色检测各浓度组成骨细胞分化水平,通过实时荧光定量PCR和免疫蛋白印迹(Western blot)法检测各浓度组的成骨细胞标记性基因及蛋白的表达差异。结果在分别干预1、2、3 d后,T3随着浓度增高,对细胞的增殖水平均呈明显上调;而且相同剂量的T3对于细胞增殖呈时间依赖性增强。此外,MC3T3-E1 Subclone 14细胞诱导7 d后,对比于0对照组,低浓度T3(1 nmol/L)对成骨细胞各分化参数均无明显作用,而10、100、1 000 nmol/L的T3对细胞ALP染色,成骨细胞相关基因及蛋白均明显上调,其中以10和100 nmol/L组最为显著。结论 T3对于MC3T3-E1 Subclone 14细胞的增殖和成骨细胞分化均有显著的促进作用。更多还原

【Abstract】 Objective To observe the effects of different concentrations of 3,3 ’,5-triiodo-L-thyronine( T3) on the proliferation and differentiation of MC3 T3-E1 subclone 14 cells. Methods Different concentrations of T3 were given to intervene cells. After 1,2 and 3 days of culture,the proliferation level of cells was measured by CCK8 method. After 7 days of intervention, the differentiation level of osteoblasts in each concentration group was detected by alkaline phosphatase( ALP) staining combined with the above-mentioned T3 administration. The difference in expression of osteoblast marker genes and proteins in each concentration group was detected by real-time fluorescence quantitative PCR and Western blotting. Results After 1,2 and 3 days of intervention, the level of cell proliferation increased significantly with the increase of T3 concentration. T3 increased the cell proliferation in a timedependent manner. In addition,7 days after MC3 T3-E1 subclone 14 cells were induced,compared with the control group of 0 nm,low concentration T3( 1 nmol/L) had no significant effects on the differentiation parameters of osteoblasts,while 10,100,and 1 000 nmol/L T3 had significant up regulation on the ALP staining of osteoblasts,among which 10 and 100 nmol/L were the most significant. Conclusion T3 significantly promotes the proliferation of MC3 T3-E1 subclone 14 cells and differentiation of osteoblasts.更多还原