摘要
目的:了解成骨细胞凋亡发生机制,观察肿瘤坏死因子-α(TNF-α)诱导大鼠成骨细胞凋亡过程中Bcl-2,Bax蛋白表达变化及意义。方法:以原代培养的SD大鼠颅骨成骨细胞为实验对象,分别用含量为0,10,20,30,40μg/LTNF-α刺激成骨细胞。通过对细胞活性、超微结构变化观察TNF-α对成骨细胞凋亡的诱导作用;同时检测细胞Bcl-2,Bax蛋白表达变化。结果:随刺激含量的加大,成骨细胞的活性逐渐降低,细胞凋亡逐渐增加。TNF-α含量为<30μg/L时Bax表达呈稳步上升趋势,30～40μg/L时开始下降。Bcl-2表达在含量为10μg/L的时候轻度上升,在20μg/L回复到正常,后随着剂量的加大Bcl-2表达开始明显下降,30～40μg/L时下降非常显著。对于Bax/Bcl-2相对表达强度与细胞死亡率做相关性分析两者呈显著性正相关(n=25,r=0.827,P<0.001)。结论:蛋白相对比例是决定成骨细胞对TNF-α诱导凋亡的易患性的重要因素。
AIM: To explore the expression and significance of Bcl 2 and bax protein in tumor necrosis factor α (TNF α ) induced apoptosis in rat osteoblasts and the mechnism of apoptosis in osteoblasts. METHODS:Primary cultured calvaria osteoblasts of rats were incited with TNF α of 0, 10, 20, 30, 40 μ g/L for 24 h.Cell activity stain and ultrastructure were used to determine the condition of osteoblastic cell apoptosis. The expressions of Bcl 2 and Bax proteins were examined by using immunochemical staining combined with imaging analysis process.RESULTS:Survival of osteoblasts decreased and apoptosis increased in dose dependent manner. TNF α had the strong effect to promote the expression of bax dose dependently when dose was 0- 30 μ g/L and depress it at 30- 40 μ g/L. Meanwhile, Bcl 2 expression slightly rose at concentration of 10 μ g/L and recovered at 20 μ g/L, and then decreased significantly at 30- 40 μ g/L. There was a significantly positive relationship between the death rate and the expression intensity of Bax/Bcl 2( n=25, r= 0.827, P< 0.001) .
出处
《中国临床康复》
CSCD
2003年第23期3210-3211,共2页
Chinese Journal of Clinical Rehabilitation
