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背景:机械应力在成骨细胞功能调控中的作用是近年骨组织生物力学研究的一个重点。流体剪切应力是否可以诱导成骨细胞内核结合因子α1的表达,其规律如何?目前少见报道。目的:观察流体剪切应力对骨肉瘤成骨样细胞(MG-63)核结合因子α1基因表达的作用。设计:对比观察实验。单位:解放军第四军医大学航空航天医学系航空航天生物动力学教研室完成。材料:MG-63人源骨肉瘤成骨样细胞。方法:实验于2004-11/2005-04在解放军第四军医大学航空航天医学系航空航天生物动力学教研室完成。①对MG-63细胞进行培养,传代60h后,将细胞置于流室系统中进行刺激实验。流体剪切应力分别为0.5Pa(0.5Pa应力刺激组)和1.5Pa(1.5Pa应力刺激组),作用时间分别为15,30和60min。同时取盖玻片置于含培养基的培养皿中,作为流体剪切应力刺激组的即时对照(对照组)。②提取细胞总RNA,进行反转录聚合酶链反应,检测细胞内核结合因子α1mRNA的表达,并计算与内参照GAPDHmRNA的比值。主要观察指标:不同流体剪切应力及不同作用时间核结合因子α1mRNA的表达值。结果:①与对照组相比,流体剪切应力刺激组的核结合因子α1mRNA的表达在作用30min和60min均显著增强,且在一定的时间范围内(15～60min),核结合因子α1mRNA的表达水平随着作用时间的延长、应力水平的增加而增强。②1.5Pa应力刺激组在作用30min和60min核结合因子α1mRNA的表达比0.5Pa应力刺激组均显著增强(P<0.01)。结论:流体剪切应力可以促进成骨细胞内核结合因子α1的表达。
BACKGROUND: The role of mechanical stress in the regulation of osteoblast function is a focus of biomechanical study of bone tissue in recent years. Whether the fluid shear stress can induce the expression of osteonuclear factor-1, and how its rules? Objective: To observe the effect of fluid shear stress on the expression of nuclear factor-1 (α1) gene in osteosarcoma osteoblast-like cells (MG-63). Design: Contrast observation experiment. Unit: PLA Fourth Military Medical University Aerospace Department of Aerospace Biodynamics Department completed. Materials: MG-63 human osteosarcoma osteoblast-like cells. METHODS: The experiment was performed at the Department of Aerospace Biodynamics, Department of Aeronautics and Astronautics, Fourth Military Medical University, PLA from November 2004 to April 2005. ① MG-63 cells were cultured, passage 60h, the cells were placed in the flow chamber system to stimulate the experiment. Fluid shear stress was 0.5Pa (0.5Pa stress stimulation group) and 1.5Pa (1.5Pa stress stimulation group), the action time were 15,30 and 60min. At the same time, coverslips were placed in media-containing Petri dishes as an immediate control of the fluid shear stress stimulus group (control group). ② Total RNA was extracted and reverse transcriptase-polymerase chain reaction was performed to detect the expression of nuclear factor-α1 mRNA. The ratio of GAPDH mRNA to internal reference was calculated. MAIN OUTCOME MEASURES: Expression values of nuclear binding factor α1 mRNA under different fluid shear stress and different time of action. Results: ①Compared with the control group, the expression of nuclear factor α1 mRNA in fluid shear stress group increased significantly at 30min and 60min, and at a certain time (15 ~ 60min), the expression of nuclear factor α1mRNA With the extension of the role of time, increased stress levels increased. ② The expression of nuclear factor-α1 mRNA in 1.5Pa stressed group was significantly increased at 30min and 60min (P <0.01) than that in 0.5Pa stress group. CONCLUSION: Fluid shear stress can promote the expression of nuclear factor-1 in the osteoblasts.