小鼠成骨细胞Wnt信号通路相关因子表达与胰岛素样生长因子1的影响

背景:胰岛素样生长因子1可促进细胞增殖、分化,但其具体作用机制尚不清楚。目的:观察胰岛素样生长因子1对小鼠成骨细胞增殖﹑分化的影响,及Wnt信号通路相关因子mRNA的表达。方法:向体外培养的小鼠成骨细胞中加入25μg/L的胰岛素样生长因子1,分别于培养的第1,2,3,4,5天用CCK-8比色法检测细胞增殖率;于培养的第3,6,9天应用酶联免疫法检测细胞内碱性磷酸酶活性;细胞培养第3天提取总RNA,采用Real-timeRT-PCR检测Wnt-3a,低密度脂蛋白受体相关蛋白5,β-catenin mRNA的表达。结果与结论:25μg/L胰岛素样生长因子1能够促进成骨细胞增殖,提高碱性磷酸酶活性,而且明显增加成骨细胞中Wnt-3a、低密度脂蛋白受体相关蛋白5、β-catenin mRNA的表达(P<0.05)。说明胰岛素样生长因子1能够促进成骨细胞的增殖和分化,Wnt信号通路参与了该调控过程。

Effects of insulin-like growth factor 1 on expression of Wnt signaling pathway-related factors in mouse osteoblasts

BACKGROUND:Insulin-like growth factor 1 (IGF-1) plays a crucial role in cell occurrence and development by regulating cell proliferation, differentiation and inhibition of apoptosis. But the underlying mechanism is unclear. OBJECTIVE:To investigate the effects of IGF-1 on the proliferation and differentiation of the cultured mouse osteoblasts and the mRNA expression levels of some related factors of Wnt signaling pathway. METHODS:The medium containing 25 μg/L IGF-1 was added to the mouse osteoblasts cultured in vitro. Cell proliferation was determined using CCK-8 method on days 1, 2, 3, 4, and 5 of culture. Alkaline phosphatase activity was determined with enzyme-linked immunosorbent assay on days 3, 6 and 9 days of culture. On day 3 of culture, total RNA was extracted, and the mRNA expression levels of Wnt-3a, low density lipoprotein receptor-related protein 5 and β-catenin were detected by real time polymerase chain reaction. RESULTS AND CONCLUSION: 25 μg/L IGF-1 can promote the proliferation of mouse osteoblasts, increase the activity of alkaline phosphatase, and significantly increase the mRNA expression levels of Wnt-3a, low density lipoprotein receptor-related protein 5 and β-catenin (P < 0.05). These findings suggest that IGF-1 can promote the proliferation and differentiation of osteoblasts and Wnt signaling pathway is involved in this process.