基础医学与临床 ›› 2013, Vol. 33 ›› Issue (11): 1404-1409.

• 间充质干细胞与皮肤损伤修复专题 • 上一篇    下一篇

EGF促进人羊膜间充质干细胞迁移上调MMP14和IL-1β表达

谭丽萍1,庞希宁1,施萍2,王竟3,王喜良1   

  1. 1. 中国医科大学
    2. 中国医科大学附属第一医院
    3. 中国医科大学附属第一医院肛肠外科
  • 收稿日期:2013-08-26 修回日期:2013-09-17 出版日期:2013-11-05 发布日期:2013-10-28
  • 通讯作者: 庞希宁 E-mail:pangxining@126.com
  • 基金资助:
    国家重点基础研究发展计划;国家自然科学基金;辽宁省科学技术计划;沈阳市科学技术计划

EGF promotes migration and expressions of MMP14 and IL-1β in human amniotic mesenchymal stem cells

  • Received:2013-08-26 Revised:2013-09-17 Online:2013-11-05 Published:2013-10-28

摘要: 目的 研究表皮细胞生长因子(EGF)促进人羊膜间充质干细胞(hAMSCs)迁移过程对基质金属蛋白酶14(MMP14)和白细胞介素1β(IL-1β)表达的影响 探讨EGF促进hAMSCs迁移的机制 方法 贴壁法分离培养原代细胞流式细胞术鉴定Transwell小室检测EGF对hAMSCs迁移能力影响;Western blot和 Real-time PCR检测MMP14和IL-1β蛋白及mRNA表达 结果 原代细胞表达间充质干细胞表面标记CD44,不表达CD45;不同浓度EGF作用hAMSCs 24h 100μg/LEGF迁移指数最大为2.0;100μg/LEGF作用hAMSCs12、24和48h迁移指数为1.6、2.0和1.7; MMP14蛋白表达12h为(0.81±0.20)显著高于对照组的(0.63±0.12)(P<0.05) ; IL-1β蛋白表达48h为(0.55±0.16)显著高于对照组的(0.30±0.05)(P<0.05) MMP14和IL-1βmRNA表达上调显著高于对照组(P<0.05) 结论 EGF促进hAMSCs迁移 上调MMP14和IL-1β表达;MMP14、IL-1β和EGF协同激活MAPK/ERK、P13K/AKT信号通路参与调控EGF促进hAMSCs迁移。

关键词: 基质金属蛋白酶14, 白细胞介素1β, 表皮生长因子, 羊膜间充质干细胞, 细胞迁移

Abstract: Objective Research the expression regulation of epidermal growth factor (EGF) on matrix metalloproteinase 14 and interleukin-1β in the progress of human amniotic mesenchymal stem cell migration .Explore the mechanisms of EGF to promote hAMSCs migration Method Take health cesarean cultured amniotic membranes adherent mesenchymal stem cells, Using Transwell cell migration in vitro system of observation EGF on migration of hAMSCs ;By Western blotting (western blot) detection EGF action hAMSCs different time MMP14 and IL-1β protein expression changes,Using real-time quantitative PCR (Real-time PCR) detection of EGF action hAMSCs different time MMP14 and IL-1β expression levels changes of mRNA Results Primary isolated and cultured cells of human amniotic cells Identified by flow cytometry express surface marker of mesenchymal stem cells CD44 do not express CD45,Transwell chamber detect different concentration of EGF acts on hAMSCs 24h reveal that 100μg/LEGF is the best:2.0,100μg/L EGF acts on hAMSCs12h、24h and 48h,migration index namely1.6、2.0 and 1.7 ;The protein level of MMP14 and IL-1β were markedly augmentmented by 100ng/mi EGF 12h results of MMP14 are(0.81±0.20)significantly higher than control group(0.63±0.12);the 48h results of IL-1β are(0.55±0.16)significantly higher than control group(0.30±0.05), 100μg/L EGF upregulated the express of MMP14 and IL-1β mRNA significantly higher than control group(P<0.05) ; Conclusion EGF have a significant role in promoting the migration of hAMSCs,in this progess EGF also upregulated the express of MMP14 and IL-1β. MMP14 IL-1β and EGF can synergistically activate the MAPK / ERK and P13K/AKT signaling pathway and MMP14 and IL-1β are involved in the regulation of EGF to promote hAMSCs migration.

Key words: matrix metalloproteinase14, interleukin 1β, epidermal growth factor, amniotic mesenchymal stem cells, cell migration

中图分类号: