中文摘要：

目的：探讨肿瘤坏死因子-α（TNF-α）对成骨-破骨细胞共育体系中成骨细胞（OB）凋亡的影响及益骨胶囊含药血清的保护作用。方法：（1）将20只10月龄SD大鼠随机分为益骨胶囊灌胃组和生理盐水灌胃组，制备含药血清与空白血清；（2）将分离的OB与破骨细胞（OC）分别接种到共育体系中，加培养液培养后，同时添入20μg/L、30μg/L、40μg/L、50μg/L、60μg/L TNF-α分别作用24h、48h、72h诱导OB凋亡，用DNA电泳法确定最佳诱导方案；（3）将细胞分为TNF-α组、TNF-α＋含药血清组、TNF-α＋空白血清组和正常对照组，分别加入DMEM培养液、含药血清培养液、空白血清培养液、DMEM培养液，除正常对照组加入PBS外，其它各组均同时加入60μg/L TNF-α，培养72h后，用荧光染色法、DNA电泳法、流式细胞仪观察各组OB凋亡情况。结果：（1）60μg/L TNF-α作用72h后，共育体系中OBDNA电泳呈较典型的梯形改变；（2）TNF-α＋含药血清组中0BDNA电泳仅出现二个条带，荧光染色可见大多数细胞均匀染色，其凋亡率（9．60％±0．26％）明显低于TNF-α组（26．90％±0．06％）与TNF-α＋空白血清组（18．10％±0．06％），P〈0.01。结论：60μg/L TNF-α作用72h可诱导共育体系中OB凋亡；益骨胶囊含药血清对TNF-a诱导的OB凋亡具有抑制作用。

英文摘要：

AIM： To observe the effect of TNF-α and Yigu capsule drug-containing sera on osteoblasts ap-optosis in osteoblasts - osteoclasts cocuhure system. METHODS： （1） Twenty female Sprague - Dawley rats, 10 months old, were randomly assigned into 2 groups, NS and Yigu capsule groups, to prepare blank sera and drug-containing sera. （2） The DNA gel electrophoresis method was used to detect apoptosis of osteoblasts treated with different concentration of TNF-α in order to determine the best dosage in the co-culture system. （3） The cells were divided into four groups, TNF-α group, normal group, TNF-α ＋ blank serum group and TNF-α ＋ drug - containing serum group. DNA gel elec-trophoresis, acridine orange staining and flow cytometry were used to observe osteoblast apoptosis in these groups. RESULTS： （1） After induced by TNF-α for 72 h at a concentration of 60 p,g/L, relatively typical DNA ladder appeared in TNF-α group. （2） Only two DNA brands appeared and most of cells were well -proportioned stained in TNF-α ＋ drug -containing serum group, the rate of osteoblasts apoptosis in TNF-α ＋ drug - containing serum group （ 9.60%±0.26%） was obviously lower than that in the TNF-α group （ 26.90%±0. 06% ） and TNF-α ＋ blank serum group （18. 10%±0.06% ）. CONCLUSION： TNF-α induces osteoblast apoptosis in the co-culture system, and Yigu capsule drug- containing serum prevents osteoblast apoptosis induced by TNF-α.