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脉冲电磁场对大鼠骨髓间充质干细胞增殖、成骨分化和Wnt/β-catenin信号通路的影响

Effect of Pulsed Electromagnetic Fields on Osteogenic Differentiation and Wnt/β-catenin Signaling Pathway in RatBone Marrow Mesenchymal Stem Cells

作者:周予婧, 王朴, 陈红英等

Author:ZHOU Yu-jing, WANG Pu, CHEN Hong-ying. et al

收稿日期:          年卷(期)页码:2015,46(3):347-353

期刊名称:四川大学学报(医学版)

Journal Name:JOURNAL OF SICHUAN UNIVERSITY (MEDICAL SCIENCE EDITION)

关键字:脉冲电磁场 骨髓间充质干细胞 成骨分化 Wnt/β-catenin信号通路

Key words:Pulsed electromagnetic fields Bonemarrow mesenchymal stem cells Osteogenic differentiation Wnt/β-catenin signaling pathway

基金项目:

中文摘要

目的 探讨脉冲电磁场(pulsed electromagnetic fields, PEMFs)对大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells, BMSCs)增殖、成骨分化以及对Wnt/β-catenin信号通路的影响。方法 将培养的第3代大鼠BMSCs分为3组。3个组均以LG-DMEM完全培养基培养1 d,待细胞贴壁后更换新鲜培养基,对照组不做干预处理; PEMFs组进行8 Hz、3.8 mT的PEMFs干预,每天40 min,持续21 d;成骨诱导基(OM)组加入成骨诱导剂,不予磁场干预。采用MTT法测定增殖活性;碱性磷酸酶(ALP)、茜素红S染色进行成骨分化鉴定;利用实时荧光定量PCR(RT-PCR)技术检测Wnt/β-catenin信号通路及成骨分化相关基因的表达。结果 相较对照组,成骨诱导剂在干预第7、14、21 d可提高BMSCs的增殖活性(PPWnt1、Wnt3a、LRP5、β-catenin、BMP-2、Runx2、ALP、OC mRNA的表达相较于对照组在相应时间点均有所上调(P

英文摘要

Objective To investigate the effect of pulsed electromagnetic fields (PEMFs) on osteogenic differentiation and Wnt/β-catenin signaling pathway in rat bone marrow mesenchymal stem cells (BMSCs). Methods Rat BMSCs were isolated and the passage 3 cells were divided into 3 groups. Cells were cultured in LG-DMEM complete medium for 1 d to ensure fully adherent. Then, change the medium. Cells were maintained in complete medium (Control group) or in osteo-induction medium (OM group). The cells in PEMFs group were cultured in complete medium and exposed to 8 Hz, 3.8 mT PEMF stimulation for 40 min/d. The intervention lasted for 21 d. Cell proliferation activity was determined by using MTT. The effects of PEMF onosteogenic differentiation were assessed by ALP and Alizarin Red S staining. Various osteoblast-relevant genes and genes of Wnt/β-catenin signaling were analyzed by quantitative real-time RT-PCR. Results We found that OM could significantly promote the proliferation of BMSC at 7 d, 14 d, 21 d (PPWnt1, Wnt3a, LRP5, β-catenin, BMP-2, Runx2, ALP, OC at special time point (P

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