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 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 DMEMmedium 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-17 F in the culture medium : 0 ng/m L group(control group),1 ng/m L group,10 ng/m L group,20 ng/m L group,50 ng/m L group,and 100 ng/m L group. After3 days,the proliferation rate of osteoblasts was detected with CCK-8 kit. The m RNA transcription and protein expressions of Runx2 and Osterix were detected with real time fluorescent quantitative PCR and Western blotting,respectively.After 10 days,the mineralized nodules were stained with Alizarin red in the control group and 100 ng/m L group. Results The cell proliferation rate was higher in 20 ng/m L,50 ng/m L and 100 ng/m L groups than in the control group( P 〈0. 05),in a concentration-dependent manner. The levels of m RNA transcription and protein expressions of Runx2 and Osterix in 50 ng/m L group and 100 ng/m L group were higher than those in the control group( P 〈 0. 05). The positivestaining area of mineralized nodules in 100 ng/m L group was larger than that in the control group. Conclusion Interleukin-17 F can promote the osteogenesis of rat osteoblasts in vitro.
出处
《山东大学学报(医学版)》
CAS
北大核心
2017年第8期24-29,共6页
Journal of Shandong University:Health Sciences
基金
山东省自然科学基金(ZR2014HM055)