Abstract: Objective To investigate the effects of vitamin D₃ (VitD₃) on osteoblasts in rats under hypoxia. Methods Newborn rat jaw osteoblasts were extracted and treated with different concentrations of cobalt chloride (0 nmol/mL, 100 nmol/mL, 200 nmol/mL, 300 nmol/mL). The proliferation of osteoblasts at 1, 3 and 5 days was detected by CCK-8 method. Flow cytometry was used to detect the apoptosis rate of osteoblasts on day 3. Osteoblasts were randomly divided into blank control group (0 nmol/mL CoCl₂), CoCl₂ group (300 nmol/mL CoCl₂) and VitD₃ group (300 nmol/mL CoCl₂ + 1 nmol/L VitD₃). VitD₃ group 2(300 nmol/mL CoCl₂ + 5 nmol/L VitD₃). The proliferative changes of osteoblasts at day 1, 3 and 5 under hypoxia were detected by CCK-8 method, and the expression of genes related to osteogenic differentiation of osteoblasts at day 10 under hypoxia was detected by qRT-PCR. Result sCompared with the control group, 200 nmol/mL CoCl₂ and 300 nmol/mL CoCl₂ had obvious inhibitory effect on osteoblast proliferation, and 300 nmol/mL CoCl₂ group had more obvious inhibitory effect on osteoblast proliferation. CoCl₂ promoted the apoptosis of osteoblasts in all groups, and 300 μmol/L was the most obvious. The apoptosis rate of osteoblasts increased with the increase of CoCl₂ concentration. Both 1 nmol/L and 5 nmol/L VitD₃ showed promoting effect on osteoblast proliferation under hypoxia condition, and the effect of 1 nmol/L VitD₃ was more significant. In terms of improving hypoxia and promoting osteoblast differentiation, 5 nmol/L VitD₃ showed superior promoting effect. Conclusion VitD₃ can promote the proliferation and differentiation of osteoblasts in rats under hypoxia.

Key words: vitamin D₃, hypoxia, osteoblasts, proliferation, osteogenic differentiation

摘要： 目的 探讨维生素D₃对缺氧状态下大鼠成骨细胞的影响。方法 提取新生大鼠颌骨成骨细胞,用不同浓度氯化钴（0 nmol/mL,100 nmol/mL,200 nmol/mL,300 nmol/mL）作用于成骨细胞,CCK-8法检测1、3、5 d成骨细胞增殖变化;流式细胞技术检测成骨细胞第3d凋亡率的变化。将成骨细胞随机分成空白对照组（0 nmol/mL CoCl₂）、CoCl₂组（300 nmol/mL CoCl₂）、VitD₃组1（300 nmol/mL CoCl₂ +1 nmol/L VitD₃）,VitD₃组2（300 nmol/mL CoCl₂ +5 nmol/L VitD₃）。CCK-8法检测缺氧状态下成骨细胞1、3、5 d增殖变化,qRT-PCR检测缺氧状态下第10 d成骨细胞成骨分化相关基因的表达。结果 与对照组比较,200 nmol/mL CoCl₂和300 nmol/mL CoCl₂对成骨细胞增殖有明显的抑制作用,其中300 nmol/mL CoCl₂组对细胞的抑制更为明显;各实验组CoCl₂对成骨细胞凋亡均有促进作用,其中以300 μmol/L最为明显,成骨细胞的凋亡率随着CoCl₂的浓度增加而增加;1 nmol/L和5 nmol/L的VitD₃均对缺氧状态下的成骨细胞增殖表现出了促进作用,1 nmol/L的VitD₃效果更为显著;在改善缺氧状态促进成骨细胞分化方面,5 nmol/L VitD₃则表现出了更为优越的促进作用。结论 维生素D₃可以促进缺氧状态下大鼠成骨细胞的增殖分化。

关键词: 维生素D₃, 缺氧, 成骨细胞, 增殖, 成骨分化