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目的:研究强磁重力环境对MG63成骨样细胞钙离子浓度和钙离子下游信号分子表达的影响。方法:利用大梯度强磁场提供μg(12T),1g(16T)和2g(12T)三组不同强磁重力复合环境处理MG63成骨样细胞后,经Fluo-3/AM标记的细胞用激光共聚焦显微镜检测处理0.5 h对细胞胞内游离钙离子浓度([Ca2+]i)的影响;用Western blot检测处理3 h对钙调蛋白(CaM)和肌球蛋白轻链激酶(MLCK)表达以及钙离子/钙调蛋白依赖蛋白激酶Ⅱ(CaMKⅡ)活性的变化。结果:钙离子浓度检测结果表明,与对照组细胞(1 g,地磁)相比,1 g(16 T)组细胞Fluo-3荧光强度增加,结果显示强磁场导致[Ca2+]i增加;与2 g(12 T)组相比,μg(12 T)组细胞Fluo-3荧光强度下降,结果显示模拟失重导致[Ca2+]i降低,抑制钙离子信号。蛋白质表达的检测结果表明,与对照组相比,1 g组细胞CaM和MLCK表达以及CaMKⅡ活性没有明显变化;与2 g(12 T)组相比,μg(12 T)组细胞CaM表达以及CaMK活性下降,结果显示模拟失重抑制CaM/CaMKⅡ信号。结论:强磁场导致MG63成骨样细胞胞内游离钙离子浓度增加,模拟失重抑制成骨样细胞钙离子/钙调蛋白信号。
OBJECTIVE: To study the effect of the magnetic environment on the expression of calcium ion and calcium ion in MG63 osteoblast-like cells. Methods: The MG63 osteoblast-like cells were treated with three different groups under the conditions of 1 g (12T), 1g (16T) and 2g (12T) in a large gradient magnetic field. Fluorescein- The effect of 0.5 h treatment on intracellular free calcium concentration ([Ca 2+] i) was detected by confocal microscopy. The expressions of CaM, MLCK, Changes of ion / calmodulin - dependent protein kinase Ⅱ (CaMK Ⅱ) activity. Results: The results of calcium ion concentration test showed that the Fluo-3 fluorescence intensity of cells in 1 g (16 T) group increased compared with control group cells (1 g, geomagnetic), and the results showed that the strong magnetic field resulted in the increase of [Ca2 +] i; Fluorescence intensity of Fluo-3 in 12 μg T (12 T) group decreased compared with that in control group (12 T). The results showed that simulated weightlessness decreased [Ca 2+] i and inhibited calcium signal. The results of protein expression showed that the expression of CaM and MLCK and the activity of CaMKⅡ in 1 g group had no significant change compared with the control group. Compared with 2 g (12 T) group, the expression of CaM and CaMK Activity decreased, the results showed that simulated weight loss inhibited CaM / CaMK Ⅱ signal. CONCLUSION: Strong magnetic field leads to an increase in the intracellular free calcium concentration in MG63 osteoblast-like cells, and simulated weightlessness inhibits osteocalcin-like calcium / calmodulin signaling.