50 Hz 1 mT脉冲电磁场对脐带间充质干细胞免疫调节能力的影响及其安全性评价

Effect and safety evaluation of 50-Hz 1-mT pulsed electromagnetic field on the immunomodulation of human umbilical cord mesenchymal stem cells

    摘要
  • 摘要:
    目的探讨50 Hz 1 mT脉冲电磁场（PEMF）对脐带间充质干细胞（MSC）免疫调节能力的影响及其安全性。
    方法采用50 Hz 1 mT PEMF辐照脐带MSC，MTT法检测辐照后细胞活性的变化；实时荧光定量PCR法检测脐带MSC的环氧合酶2（COX2）、IL-10、吲哚胺-2，3-双加氧酶（IDO）、肝细胞生长因子（HGF）和转化生长因子β（TGF-β）等基因mRNA表达水平的改变；流式细胞仪检测PEMF对脐带MSC活性氧（ROS）水平、细胞凋亡和细胞周期的影响；单细胞凝胶电泳分析脐带MSC DNA损伤的情况。采用SPSS19.0软件，对各组间数据进行独立样本t检验分析。
    结果50 Hz 1 mT PEMF辐照可增强脐带MSC的细胞活性，其中每天辐照4 h组变化最为明显，差异有统计学意义（t=3.505，P < 0.05）。PEMF辐照可促进COX2、HGF、IDO和TGF-β等免疫调节因子mRNA的表达，但变化不明显，差异无统计学意义（t=0.982~2.376，均P>0.05），对于干扰素γ（IFN-γ）激活的MSC，PEMF辐照可显著上调COX2、HGF、IDO和TGF-β的mRNA表达水平，差异有统计学意义（t=2.436~3.747，均P < 0.05）。50 Hz 1 mT PEMF辐照对脐带MSC的ROS水平、DNA损伤及细胞凋亡率、细胞周期无显著影响。
    结论对于IFN-γ激活的MSC，50 Hz 1 mT PEMF可显著上调COX2、HGF、IDO和TGF-β的mRNA表达水平，从而增强脐带MSC的免疫调节能力。50 Hz 1 mT PEMF对脐带MSC的ROS水平、细胞凋亡及DNA损伤无明显影响，此参数下的PEMF辐照对脐带MSC是安全的。

     

    Abstract:
    ObjectiveTo investigate the effect of 50-Hz, 1-mT pulsed electromagnetic field (PEMF) on the immunomodulation of human umbilical cord mesenchymal stem cells(MSCs) and to evaluate the safety of PEMF as a therapeutic strategy.
    MethodsChanges in cell viability was observed via MTT. Real-time PCR was used to detect the expression levels of cyclooxygenase 2(COX2), indoleamine 2, 3-dioxygenase(IDO), IL-10, hepatocyte growth factor(HGF), and transforming growth factor-β(TGF-β) in MSCs. To explore the safety of PEMF, a flow cytometry experiment was conducted to observe apoptosis and reactive oxygen species(ROS) level. DNA damage was detected via comet assay. Data were statistically analyzed via t-test with SPSS 19.0. P < 0.05 was considered statistically significant.
    ResultsCell viability increased, with the most obvious increase in 4 h/d(t=3.505, P < 0.05). In interferon-γ(INF-γ)-activated MSC, the expression levels of immunomodulation factors improved after PEMF irradiation(t=2.436-3.747, all P < 0.05). Furthermore, PEMF did not induce cell apoptosis, ROS generation, and DNA damage.
    Conclusion50-Hz, 1-mT PEMF improves MSC immunomodulation and may not cause radiation injury.

     

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