机械离心力对成骨细胞骨形态发生蛋白信号通路中Runx-2 mRNA的影响  

Effect of mechanical centrifugal force on Runx-2 mRNA in osteoblasts' bone morphogenetic protein signal pathway

作　　者：段峰[1] 关键[1] 杨红岩[2] 王心彧[1] 张国梁[1] 朱杨[1] 

机构地区：[1]佳木斯大学附属口腔医院,黑龙江省佳木斯市154004 [2]深圳市龙岗区横岗人民医院口腔科,广东省深圳市518115

出　　处：《中国组织工程研究》2014年第33期5305-5309,共5页Chinese Journal of Tissue Engineering Research

基　　金：黑龙江省教育厅自然科学面上项目(11551493)~~

摘　　要：背景:机械力对成骨细胞生理活动存在一定影响,Runx-2是骨形态发生蛋白信号的靶目标,是调节成骨细胞分化的重要因子,骨形态发生蛋白信号转导通路参与了成骨细胞对机械离心力刺激的生理响应过程。目的:观察不同转速及时间作用下,机械离心力对成骨细胞骨形态发生蛋白信号通路的影响。方法:将MC3T3-E1细胞用含体积分数10%胎牛血清的DMEM培养基预处理24 h后,分为对照组,90 r/min组、180 r/min及250 r/min组,每组再分为离心6,12,24 h亚组,给予不同转速离心力和相同转速不同时间的刺激,重复实验3次。对照组同步置于除离心外相同的环境中。提取总RNA,并反转录为cDNA,通过实时荧光定量PCR检测Runx2基因表达情况。结果与结论:随着加力时间的延长,Runx2 mRNA的表达增加,二者成正相关,转速为180 r/min组的Runx2mRNA表达明显高于90 r/min组和250 r/min组(P<0.01);90 r/min组和250 r/min组Runx2 mRNA的表达均高于对照组(P=0.039),且都随时间延长差异明显。结果可见离心力大小和离心持续时间不同对成骨细胞骨形态发生蛋白信号通路的生理响应不同,该通路在对力学信号引起的信息传递级联反应中起重要作用。BACKGROUND：Mechanical strain certainly has an effect on physiological activities of osteoblasts. Runx-2 is a target of bone morphogenic protein signal and is an important factor for regulation of osteoblastic differentiation. Bone morphogenic protein signal transduction pathway is involved in physiological response of osteoblast to stimulation of mechanical centrifugal force. OBJECTIVE：To observe the effect of mechanical centrifugal force on bone morphogenetic protein signal pathway under different time period and speed. METHODS：MC3T1-E1 cells were pre-treated in DMEM medium containing 10%fetal bovine serum for 24 hours, and then divided into control group, 90 r/min group, 180 r/min group and 250 r/min group. Each group was then subdivided into 6 hours, 12 hours and 24 hours centrifugation subgroups. Experiments were repeated for three times for different centrifugal speed and different time period. Except centrifugation, the control group was under the same environment. Total RNA was extracted and reversely transcribed into cDNA. Runx-2 gene expression was determined by real time fluorescent quantitative PCR. RESULTS AND CONCLUSION：The expression of Runx-2 mRNA was increased with extension of time, showing a positive correlation between the two. The mRNA expression at 180 r/min was significantly higher than that at 90 r/min and 250 r/min （P〈0.01）;at 90 r/min and 180 r/min, the Runx-2 mRNA expression was higher than that in the control group （P=0.039）, both of them showed significant difference along with the time. The difference of centrifugal force speed and duration is associated with different physiological response of osteoblasts in bone morphogenic protein signal pathway, which plays an important role in mechanical signal transmission and cascade reaction.

关 键 词：组织构建 骨组织工程 机械离心 成骨细胞 信号通路 骨形态发生蛋白 RUNX2 MRNA 

分 类 号：R318[医药卫生—生物医学工程]