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目的:研究模拟微重力对前成骨细胞的增殖作用及其分子机制。方法:利用2D回转器模拟失重条件培养前成骨细胞MC3T3-E1 24小时;将p-AKT激活剂SC79加入细胞培养基后模拟失重条件下培养前成骨细胞MC3T3-E1 24小时;利用Western blot技术分别检测细胞增殖相关蛋白PCNA以及AKT、p-AKT的表达变化情况。结果:(1)与对照组相比,模拟失重组前成骨细胞增殖受到抑制(P<0.01),p-AKT/AKT比值减小(P<0.01);(2)加入SC79组与对照组相比,p-AKT/AKT比值显著增加;(3)加入SC79的模拟失重组(MG+SC79)与模拟失重组相比(MG),p-AKT/AKT比值增加,PCNA蛋白表达增加(P<0.01),成骨细胞增殖有所恢复。结论:模拟微重力可能通过抑制AKT的磷酸化形式抑制前成骨细胞的增殖,加入p-AKT激活剂可部分恢复前成骨细胞的增殖。
Objective: To study the proliferation of pre-cultured osteoblasts by simulated microgravity and its molecular mechanism. Methods: The pre-cultured osteoblasts MC3T3-E1 were cultured for 24 hours in a simulated condition of weightlessness with a 2D gyrator. The pre-cultured osteoblasts MC3T3-E1 were cultured for 24 hours after adding the p-AKT activator SC79 into the cell culture medium under simulated weightlessness conditions. Technology were detected cell proliferation-related protein PCNA and AKT, p-AKT expression changes. Compared with the control group, the proliferation of osteoblasts was inhibited (P <0.01) and the ratio of p-AKT / AKT was decreased (P <0.01). (2) Compared with the control group, (3) The ratio of p-AKT / AKT was increased and the expression of PCNA was increased (P <0.05) in the simulated weightless group (MG + SC79) <0.01), osteoblast proliferation has been restored. CONCLUSIONS: Simulated microgravity may inhibit the proliferation of pre-osteoblasts by inhibiting the phosphorylation of AKT. The addition of p-AKT activator partially restored the proliferation of pre-osteoblasts.