肿瘤坏死因子α诱导人成骨细胞和人成骨细胞系HOS-8603凋亡的作用  

Apoptosis of human osteoblasts and osteoblast cell line HOS8603 induced by tumor necrosis factor alpha

作　　者：王文良 吴岳嵩[2] 杨柳[3] 许建中[3] 娄永华[4] 焦炳华[4] 

机构地区：[1]武装警察部队医学院附属医院骨科,天津市300162 [2]解放军第二军医大学长海医院骨科,上海市200433 [3]解放军第三军医大学西南医院骨科,重庆市400038 [4]解放军第二军医大学基础医学部毒理学研究室,上海市200433

出　　处：《中国临床康复》2005年第26期162-164,i0008,共4页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:观察不同浓度重组肿瘤坏死因子α对于培养的人原代成骨细胞和人成骨细胞系HOS-8603的凋亡作用,并分析浓度效应。方法:实验于1998-04/2000-04在第二军医大学卫生毒理学教研室完成。在人成骨细胞和人成骨细胞系HOS-8603中,实验组加浓度为1,10,100,1000ng/L的肿瘤坏死因子α,对照组加等量的无肿瘤坏死因子培养基,培养后检测细胞活力应用噻唑兰法,检测细胞凋亡情况应用DNA梯形条带和透射电镜等方法。结果:①肿瘤坏死因子α对培养成骨细胞和HOS-8603细胞的作用(噻唑兰法):肿瘤坏死因子α浓度为100和1000ng/L实验组均高于对照组犤成骨细胞:(0.14±0.006),(0.10±0.010),(0.27±0.013)ng/L;HOS-8603细胞:(0.15±0.012),(0.09±0.015),(0.27±0.013)ng/L,P<0.05犦。②DNA梯形条带结果:凋亡细胞使DNA在核小体外降解,形成180～200bp或与之成倍的DNA片段。在琼脂糖凝胶电泳时呈现梯形结果。在肿瘤坏死因子α浓度为1000ng/L时,成骨细胞可检测到典型DNA梯形条带。③成骨细胞的凋亡情况:应用透射电镜检查,浓度为1000ng/L实验组较浓度为100ng/L实验组凋亡细胞多。肿瘤坏死因子α同样可以诱导HOS-8603的凋亡,未加肿瘤坏死因子α时,凋亡很少,加入100ng/L及1000ng/L肿瘤坏死因子α时,凋亡细胞数量明显增加。结论:肿瘤坏死因子α质量浓度达到100g/L时,可以引起体外培养的成骨细胞和成骨细胞系HOS-8603细胞凋亡,并随浓度加大出现凋亡细胞增多的剂量-效应关系,结果验证了肿瘤坏死因子能刺激成骨细胞凋亡的假说。AIM： To investigate the influence of tumor necrosis factor alpha （TNF-α） on the apoptosis of cultured human osteoblasts and osteoblast cell line HOS-8603, and analyze the concentration-effect relationship. METHODS： The experiment was conducted in the Department of Toxicology, Second Military Medical University from April 1998 to April 2000. TNF-α at 1 , 10 , 100, 1 000 ng/L and equivalent culture medium without TNF-α were added to cultured human osteoblsts and osteoblast cell line HOS-8603 in experiment group and control group respectively. The activity of cultured cells was detected by methyl-thiazolyl-tetrazolium （MTT）. DNA ladder, transmission electron microscope and other methods were used to determine the apoptosis of cells. RESULTS： ①Effects of TNF-α on the cultured human osteoblsts and osteoblast cell line HOS-8603 measured by MTT： The concentration of osteoblsts and HOS-8603 cells in the TNF-α 100 ng/L and 1 000 ng/L experiment groups were significantly higher than that in the control group [osteoblsts： （0.14±0.006）, （0.10±0.010） ng/L vs（0.27±0.013） ng/L; HOS- 8603 cells： （0.15±0.012）, （0.09±0.015） ng/L vs（0.27±0.013） ng/L, P 〈 0.05]. ②Results of DNA ladder： The DNA was degraded outside the nucleosomes by apoptotic cells, and formed fragments of 180 to 200 bp or the fragments of geminate 180 to 200 bp. DNA was shown ladder-shaped by agarose gel electrophoresis. When the TNF-α was added at the concentration of 1 000 ng/L, typical ladder-shaped DNA was detected in the osteoblasts. ③Apoptosis of osteoblasts： Shown by transmission electron microscopy, the apoptotic osteoblasts were more in the TNF-α 1 000 ng/L group than in the TNF-α 100 ng/L group. TNF-α could also induce the apoptosis of osteoblast cell line HOS-8603. A few cells were apoptotic when no TNF-α was added, but as TNF-α at 1 000 ng/L and 100 ng/L were added, extraordinarily more cells were apoptotic. CONCLUSION： TNF-α at 100 ng/L can induce the apoptosis of osteoblasts a

关 键 词：肿瘤坏死因子 成骨细胞 细胞凋亡 HOS-8603 肿瘤坏死因子Α 人成骨细胞 凋亡作用 细胞系 成骨细胞凋亡 DNA片段 

分 类 号：R738.1[医药卫生—肿瘤] R587.1[医药卫生—临床医学]