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目的观察随机定位模拟微重力对人前破骨细胞FLG29.1增殖和分化的影响。方法 FLG29.1细胞分为正常对照组与随机定位处理组,分别培养72 h后收集细胞。采用细胞计数法检测细胞总数和活细胞数;流式细胞术检测细胞周期;Griess法检测培养基中NO(nitric oxide,NO)浓度;抗酒石酸盐酸性磷酸酶(TRAP)染色计算TRAP阳性细胞比例;对硝基苯磷酸(pNPP)法检测胞内TRAP活性。结果随机定位处理后,FLG29.1细胞增殖能力与活细胞比例均较对照组明显增加;细胞周期分布发生变化,处于G1期的细胞比例增加;培养基中NO浓度有增加趋势;在回转处理的同时添加诱导剂12-氧-十四烷酰佛波醋酸酯-13乙酸酯(TPA),发现TRAP阳性细胞数量增多;胞内TRAP活性较对照组明显增加。结论随机定位模拟微重力提高了FLG29.1细胞活力并促进其向破骨细胞分化。
Objective To observe the effects of random-positioning simulated microgravity on the proliferation and differentiation of human osteoclast FLG29.1. Methods FLG29.1 cells were divided into normal control group and random-targeted treatment group, and cultured for 72 hours respectively. The total number of cells and the number of viable cells were detected by cell counting method. The cell cycle was detected by flow cytometry. The concentration of nitric oxide (NO) was detected by Griess method. The percentage of TRAP-positive cells was calculated by TRAP staining. ; Intracellular TRAP activity was detected by pNPP method. Results After the cells were randomly positioned, the proliferation and viability of FLG29.1 cells were significantly increased compared with the control group. The cell cycle distribution was changed and the proportion of cells in G1 phase increased. The concentration of NO in the medium increased. At the same time, the inducer 12-O-tetradecanoyl phorbol-13 acetate (TPA) was added and the number of TRAP positive cells was increased. The intracellular TRAP activity was significantly increased compared with the control group. Conclusion Randomly-positioned simulated microgravity increases the viability of FLG29.1 cells and promotes osteoclast differentiation.