张力对成骨样细胞NF-κBp65表达活性的影响  

Effects on the Expression of NF-κBp65 of Osteoblast-like Cell under Stretch Load with Different Daily Loading Times

作　　者：龙洁[1] 田卫东[1] 樊瑜波[2] 冯帆[1] 李声伟[1] 

机构地区：[1]四川大学华西口腔医学院,成都610041 [2]四川大学生物力学工程实验室,成都610065

出　　处：《生物医学工程学杂志》2008年第1期88-91,共4页Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目(30070819,10502037);中国博士后科研基金资助项目(2003034420)

摘　　要：探讨采用应变值相同，但日加力次数不同的张力加载方式对成骨样细胞NF-κBp65表达的影响。将成骨样细胞株MG-63进行体外传代培养，并接种在四点弯曲细胞力学加载仪的细胞培养板上，对细胞进行张力刺激：将接种好的细胞按日加力次数的不同随机分为4组，每组4个样本，1组为阴性对照组，2～4组样本分别施以强度为2000μ strain，频率为0．5Hz的张力刺激，施力次数分别为1、2、4次／d，加力时间均为60min／d，共加力2d。各组样本均在加力后置于含10％小牛血清DMEM培养液的专用培养皿中继续培养24h，收集细胞培养板与对照组同时行NF-κBp65免疫组化染色，分别测定在力学强度相同、作用时间相同而日加力次数不同的加载条件下，成骨样细胞MG-63对NF-κBp65的表达状况并进行组间比较。结果发现各组细胞均可见NF-κBp65阳性表达，各组细胞的阳性表达率有显著性差异，各加力组细胞阳性表达率显著高于对照组，4次／d加力组细胞阳性表达率显著高于另两实验组，2次／d加力组细胞阳性表达率显著高于1次／d加力组。表明在相同强度的张力作用下，随着日施加张力次数的增加，成骨样细胞MG-63中NF-κB的表达活性逐步增强，提示在力学强度相同的条件下，采用较高日加载次数的加载手段，能更有效地提高成骨细胞的转录水平。The purpose of this study was to investigate the effect on the expression of NF-κBp65 of osteoblast-like cell under stretch load with the same amplitude but different daily loading times. The osteoblast-like cells MG-63 were passage cultured and stretched by the four-point-bend loading device; based on the daily loading times, the osteoblast-like ceils were randomly divided into four groups. The first was the control, the others were stretched with mechanical tension with the same amplitude of 2 000 μ strain and at the same frequency of 0.5 Hz., but the daily loading times were 1 time/ d, 2 times/d, 4 times/d differently for each group, the periods of mechanical tension applied to the cells of the three groups were all 60 min/d and lasted for 2d total. After the ceils being streteched, the expression levels of NF-κBp65 of the osteoblast-like cells of the three groups and control group were investigated by using the techniques of immunohistcchernistry, and were compared with each other. The results showed that the positive expression ratios of the four groups were different significantly; the positive expression ratio of the control was lower than those of the other three groups; the positive expression ratio of the 4 tirnes/d group was higher than those,e of the other two stretched groups; the positive expression ratio of the 2 times/d group was higher than that of the 1 time/d group. The results suggested that when the osteoblast-like cell was under the stretch load with different daily loading times but the same amplitude, the expression ratio of NF-κBp65 in the cell increased with the rising of the stimulating times. It means that the mechanical strain with high daily loading times could promote the transcriptional level of osteoblast-like cell more effectively.

关 键 词：张力 日加力次数 成骨样细胞 NF-ΚBP65 

分 类 号：R318.01[医药卫生—生物医学工程]