白介素17F促进大鼠成骨细胞增殖、矿化和Runx2、Osterix的表达

doi:10.6040/j.issn.1671-7554.0.2017.059

摘要/Abstract

摘要： 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wistar大鼠颅盖骨原代成骨细胞,用含10%胎牛血清的低糖DMEM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/mL组(对照组)、1 ng/mL组、10 ng/mL组、20 ng/mL组、50 ng/mL组、100 ng/mL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mRNA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/mL组进行矿化结节染色。结果 20 ng/mL组、50 ng/mL组和100 ng/mL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/mL组和100 ng/mL组成骨细胞中Runx2、Osterix的mRNA转录水平和蛋白表达水平均高于对照组(P<0.05)。100 ng/mL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。

关键词: Runx2, 白介素17F, 成骨细胞, 矿化, Osterix

Abstract: Objective To investigate the effects of interleukin-17F(IL-17F)on the proliferation, mineralization and expressions of Runx2 and Osterix in rat osteoblasts. Methods Primarycalvarial osteoblasts were isolated from neonatal Wistarrats<24 h, cultured with low glucose DMEM medium containing 10% fetal bovine serum, and the medium was replaced every 3 days. The cells were passaged when the cell fusion area reached about 80%. The fourth generation osteoblasts were divided into 6 groups according to the concentration of IL-17F in the culture medium: 0 ng/mL group(control group), 1 ng/mL group, 10 ng/mL group, 20 ng/mL group, 50 ng/mL group, and 100 ng/mL group. After 3 days, the proliferation rate of osteoblasts was detected with CCK-8 kit. The mRNA transcription and protein expressions of Runx2 and Osterix were detected with real time fluorescent quantitative PCR and Western blotting, respectively. After 10days, the mineralized nodules were stained with Alizarin red in the control group and 100 ng/mL group. Results The cell proliferation rate was higher in 20 ng/mL, 50 ng/mL and 100 ng/mL groups than in the control group(P<0.05), in a concentration-dependent manner. The levels of mRNA transcription and protein expressions of Runx2 and Osterix in 50 ng/mL group and 100 ng/mL group were higher than those in the control group(P<0.05). The positive 山东大学学报 (医学版)55卷8期 -姬原原,等.白介素17F促进大鼠成骨细胞增殖、矿化和Runx2、Osterix的表达 \=-staining area of mineralized nodules in 100 ng/mL group was larger than that in the control group. Conclusion Interleukin-17F can promote the osteogenesis of rat osteoblasts in vitro.

Key words: Runx2, Osterix, Mineralization, Interleukin-17F, Osteoblast

中图分类号:

R589

姬原原,包景泊,赵旭,曹鲁宁,崔敏,张娜,于灵芝. 白介素17F促进大鼠成骨细胞增殖、矿化和Runx2、Osterix的表达[J]. 山东大学学报（医学版）, 2017, 55(8): 24-29.

JI Yuanyuan, BAO Jingbo, ZHAO Xu, CAO Luning, CUI Min, ZHANG Na, YU Lingzhi. IL-17F promotes the proliferation, mineralization and expressions of Runx2 and Osterix in rat osteoblasts[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2017, 55(8): 24-29.

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