3,3',5-碘-L-甲腺原氨酸对成骨细胞前体增殖和成骨分化的作用

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、1000 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.

3,3',5-triiodo-L-thyronine promotes the proliferation of osteoblast precursors and differentiation of osteoblasts

Objective To observe the effects of different concentrations of 3,3 ,5-triiodo-L-thyronine(T3)on the proliferation and differentiation of MC3T3-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 ex-pression 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 time-dependent manner.In addition,7 days after MC3T3-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 osteo-blasts,while 10,100,and 1000 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 MC3T3-E1 subclone 14 cells and differentiation of osteoblasts.