文章摘要
韦盛,杨勇,赵东明,刘朝旭,吴华.白细胞介素-1β和电磁场对大鼠骨髓间充质干细胞成骨分化的影响.骨科,2019,10(4):335-339
白细胞介素-1β和电磁场对大鼠骨髓间充质干细胞成骨分化的影响
Effects of interleukin-1β on osteogenic differentiation of rat mesenchymal stem cells under electro-magnetic fields stimulation.
投稿时间:2019-03-05  
DOI:10.3969/j.issn.1674-8573.2019.04.015
中文关键词: 白细胞介素-1β  电磁场  骨髓间充质干细胞  成骨分化
英文关键词: Interleukin-1β  Electromagnetic fields  Bone mesenchymal stem cells  Osteogenic differentiation
基金项目:国家自然科学基金(51537004)
作者单位E-mail
韦盛 华中科技大学同济医学院附属同济医院骨科武汉 430030  
杨勇 华中科技大学同济医学院附属同济医院骨科武汉 430030  
赵东明 华中科技大学同济医学院附属同济医院骨科武汉 430030  
刘朝旭 华中科技大学同济医学院附属同济医院骨科武汉 430030 liu.chaoxu@tjh.tjmu.edu.cn 
吴华 华中科技大学同济医学院附属同济医院骨科武汉 430030 wuhua360@aliyun.com 
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中文摘要:
      目的 研究白细胞介素-1β(interleukin-1β, IL-1β)和低频电磁场对大鼠骨髓间充质干细胞(bone mesenchymal stem cells, BMSCs)成骨分化的影响。方法 取第三代大鼠BMSCs置于6孔板培养并随机分为4组:对照组、IL-1β组、电磁场组和IL-1β+电磁场组。IL-1β组和IL-1β+电磁场组在每次换新鲜培养基时加入1 ng/ml IL-1β,电磁场组和IL-1β+电磁场组每天给予15 Hz/1 mT的低频正弦波电磁场刺激4 h。培养3 d后提取RNA和蛋白质,用实时定量聚合酶链式反应(Real-time PCR)检测碱性磷酸酶(alkaline phosphatase,ALP)、骨桥蛋白(osteopontin, OPN)及Runt相关转录因子2(runt-related transcription factor 2, Runx2)的mRNA表达。Western blotting检测Runx2和OPN的蛋白表达。培养7 d后检测ALP、OPN及Runx2的mRNA表达。另取两组细胞,分别加入1 ng/ml的IL-1β,1 ng/ml的IL-1β+抑制剂SB203580后,给予15 Hz/1 mT的低频正弦波电磁场刺激并分别于0、5、15、30、60及120 min提取蛋白质,用Western blotting检测磷酸化p38的蛋白表达。结果 培养3 d后,IL-1β组、电磁场组和IL-1β+电磁场组中ALP、OPN的mRNA的表达量与对照组相比均有明显增高,差异均有统计学意义(P均<0.05)。与对照组相比,电磁场组和IL-1β+电磁场组中Runx2的mRNA的表达量明显升高,差异均有统计学意义(P均<0.05)。与对照组相比,IL-1β组、电磁场组和IL-1β+电磁场组中OPN、Runx2蛋白的表达量均明显增高。培养7 d后,与对照组相比,IL-1β组和IL-1β+电磁场组中ALP的mRNA表达均降低,在电磁场组则升高(P<0.05);IL-1β组、电磁场组和IL-1β+电磁场组中OPN的mRNA表达与对照组相比均增高(P均<0.05);Runx2的基因的表达量在IL-1β组及IL-1β+电磁场组较对照组均降低,差异均有统计学意义(P均<0.05)。磷酸化的p38在第30、60、120 min时的蛋白质表达较0 min时升高(P<0.05),使用p38抑制剂SB203580后磷酸化的p38表达无明显变化。结论 IL-1β和电磁场可以在早期促进大鼠BMSCs的ALP、OPN、Runx2的表达,IL-1β和电磁场可以激活磷酸化的p38。
英文摘要:
      Objective To study the effects of interleukin-1β (IL-1β) and low frequency electromagnetic fields on the osteogenic differentiation of rat mesenchymal stem cells in vitro. Methods The rat bone mesenchymal stem cells (BMSCs) of third passage were cultured in the six-well plate and randomly divided into four groups: the control group, the IL-1β group, the electromagnetic fields stimulation (EMF) group, and the IL-1β+EMF group. 1 ng/ml of IL-1β was added into the IL-1β group and the IL-1β+EMF group when each replacement of the fresh culture medium. The EMF group and the IL-1β+EMF group were treated with the low frequency sinusoidal EMF for 4 h per day. Cells were harvested on the 3rd and 7th day. Real-time PCR was used to detect the mRNA expression of alkaline phosphatase (ALP), osteopontin (OPN), and runt-related transcription factor 2 (Runx2). The expression of OPN and Runx2 proteins was detected by Western blotting. Two groups of cells were selected, one group was treated with 1 ng/ml IL-1β, and the other with 1 ng/ml IL-1β and SB203580, then 15 Hz/1 mT the low frequency sinusoidal EMF was given and the proteins were extracted separately at 0, 5, 15, 30, 60 and 120 min. Western blotting was used to detect the phosphorylated p38 protein expression. Results After 3 days of culture, the mRNA expression levels of ALP and OPN significantly increased in IL-1β group, EMF group and IL-1β+EMF group as compared with those in the control group (all P<0.05). The mRNA expression of Runx2 was significantly increased in EMF group and IL-1β+EMF group as compared with the control group (P<0.05). The protein expression of OPN and Runx2 were increased in IL-1β group, EMF group and IL-1β+EMF group as compare with the control group (all P<0.05). After 7 days of culture, the mRNA expression of ALP was significantly decreased in IL-1β group and IL-1β+EMF group, but increased in EMF group (P<0.05). The mRNA expression levels of OPN increased in IL-1β group, EMF group and IL-1β+EMF group as compared with the control group (P<0.05). The mRNA expression levels of Runx2 were decreased in IL-1β group and IL-1β+EMF group as compared with the control group (P<0.05). The expression of phosphorylated p38 protein was increased at 30, 60, and 120 min after treatment with IL-1β stimulation+EMF as compared with that at 0 min. There was no significant difference in the expression of phosphorylated p38 protein after treatment with SB203580. Conclusion IL-1β and EMF increased the expression of ALP, OPN, and Runx2 in rat BMSCs in the early stage. Phosphorylated p38 proteins could be activated by IL-1β and EMF.
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