CAJ | 학술논문

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Runx2 mRNA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。
배경：궤계력대성골세포생리활동존재일정영향，Runx-2시골형태발생단백신호적파목표，시조절성골세포분화적중요인자，골형태발생단백신호전도통로삼여료성골세포대궤계리심력자격적생리향응과정。 　　목적：관찰불동전속급시간작용하，궤계리심력대성골세포골형태발생단백신호통로적영향。 　　방법：장MC3T3-E1세포용함체적분수10%태우혈청적DMEM배양기예처리24 h후，분위대조조，90 r/min조、180 r/min급250 r/min조，매조재분위리심6，12，24 h아조，급여불동전속리심력화상동전속불동시간적자격，중복실험3차。대조조동보치우제리심외상동적배경중。제취총RNA，병반전록위cDNA，통과실시형광정량PCR검측Runx2기인표체정황。 　　결과여결론：수착가력시간적연장，Runx2 mRNA적표체증가，이자성정상관，전속위180 r/min조적Runx2 mRNA표체명현고우90 r/min조화250 r/min조(P<0.01)；90 r/min조화250 r/min조Runx2 mRNA적표체균고우대조조(P=0.039)，차도수시간연장차이명현。결과가견리심력대소화리심지속시간불동대성골세포골형태발생단백신호통로적생리향응불동，해통로재대역학신호인기적신식전체급련반응중기중요작용。
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.