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目的探讨白介素17F对大鼠成骨细胞增殖、矿化和Runx2、Osterix表达水平的影响。方法分离提取新生24 h内w istar大鼠颅盖骨原代成骨细胞,用含10%胎牛血清的低糖DM EM培养基培养,培养基每3天更换1次。当细胞相互融合生长面积达80%左右时,进行细胞传代。向培养基中加入白介素17F,根据浓度将第4代成骨细胞分为0 ng/m L组(对照组)、1 ng/m L组、10 ng/m L组、20 ng/m L组、50 ng/m L组、100 ng/m L组。分组后继续培养3 d,采用CCK-8试剂盒检测各组成骨细胞增殖率,采用实时荧光定量PCR法和Western blotting法分别检测成骨细胞中Runx2、Osterix的m RNA转录水平和蛋白表达水平。在分组培养第10天,对照组和100 ng/m L组进行矿化结节染色。结果 20 ng/m L组、50 ng/m L组和100 ng/m L组成骨细胞增殖率较对照组增高(P<0.05),呈浓度依赖性。50 ng/m L组和100 ng/m L组成骨细胞中Runx2、Osterix的m RNA转录水平和蛋白表达水平均高于对照组(P<0.05)。100 ng/m L组成骨细胞矿化结节的染色阳性区域多于对照组。结论白介素17F可促进体外培养的大鼠成骨细胞的成骨作用。
Objective To investigate the effect of interleukin 17F on proliferation, mineralization and expression of Runx2 and Osterix in rat osteoblasts. Methods Primary osteoblasts of wistar rats were isolated and extracted within 24 hours of newborn, cultured in low glucose DMEM containing 10% fetal bovine serum, and the medium was changed every 3 days. When the cells fused to each other about 80% of the growth area, cell passage. Interleukin 17F was added to the culture medium and the 4th generation osteoblasts were divided into 0 ng / m L group (control group), 1 ng / m L group, 10 ng / m L group and 20 ng / m L group , 50 ng / m L group and 100 ng / m L group. The cells were cultured for 3 days. The proliferation rate of osteoblasts was detected by CCK-8 kit. The mRNA and protein levels of Runx2 and Osterix in osteoblasts were detected by real-time fluorescence quantitative PCR and Western blotting respectively. On the 10th day of grouping, the control group and the 100 ng / m L group were subjected to mineralized nodule staining. Results The proliferation rates of osteoblasts in 20 ng / m L group, 50 ng / m L group and 100 ng / m L group were significantly higher than those in control group (P <0.05). The mRNA and protein expressions of Runx2 and Osterix in 50 ng / m L group and 100 ng / m L group were higher than those in control group (P <0.05). 100 ng / m L group osteoblast mineralized nodules were more positive staining area than the control group. Conclusion Interleukin 17F can promote the osteoblastic effect of cultured rat osteoblasts in vitro.