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IL-6诱导小鼠骨髓中性粒细胞中MMP-9的生成与释放的实验

高晗, 颜彬, 彭秋子, 刘颖华, 袁冬妮, 吴绪峰

高晗, 颜彬, 彭秋子, 刘颖华, 袁冬妮, 吴绪峰. IL-6诱导小鼠骨髓中性粒细胞中MMP-9的生成与释放的实验[J]. 肿瘤防治研究, 2016, 43(2): 129-132. DOI: 10.3971/j.issn.1000-8578.2016.02.007
引用本文: 高晗, 颜彬, 彭秋子, 刘颖华, 袁冬妮, 吴绪峰. IL-6诱导小鼠骨髓中性粒细胞中MMP-9的生成与释放的实验[J]. 肿瘤防治研究, 2016, 43(2): 129-132. DOI: 10.3971/j.issn.1000-8578.2016.02.007
GAO Han, YAN Bin, PENG Qiuzi, LIU Yinghua, YUAN Dongni, WU Xufeng. IL-6 Induces Production and Releasing of MMP-9 in Murine Bone Marrow Neutrophils[J]. Cancer Research on Prevention and Treatment, 2016, 43(2): 129-132. DOI: 10.3971/j.issn.1000-8578.2016.02.007
Citation: GAO Han, YAN Bin, PENG Qiuzi, LIU Yinghua, YUAN Dongni, WU Xufeng. IL-6 Induces Production and Releasing of MMP-9 in Murine Bone Marrow Neutrophils[J]. Cancer Research on Prevention and Treatment, 2016, 43(2): 129-132. DOI: 10.3971/j.issn.1000-8578.2016.02.007

IL-6诱导小鼠骨髓中性粒细胞中MMP-9的生成与释放的实验

基金项目: 湖北省妇幼保健院院内课题(220933002)
详细信息
    作者简介:

    高晗(1972-),男,本科,副主任医师,主要从事妇科肿瘤方面的研究

    通讯作者:

    吴绪峰,E-mail: zwuxufeng@163.com

  • 中图分类号: R737.3

IL-6 Induces Production and Releasing of MMP-9 in Murine Bone Marrow Neutrophils

  • 摘要: 目的 探讨IL-6诱导小鼠骨髓中性粒细胞(PMN)生成和释放基质金属蛋白酶-9(MMP-9)的作用及激活途径。方法 分离小鼠骨髓PMN,将其分为空白对照组、IL-6刺激组、IL-6+DMSO组、IL-6+STAT3抑制剂Ⅷ组。实时荧光定量聚合酶链式反应检测细胞内MMP-9 mRNA表达,明胶酶谱检测PMN释放的MMP-9活性,Western blot检测PMN中STAT3蛋白磷酸化改变。结果 IL-6刺激后,PMN中MMP-9 mRNA水平明显升高(P=0.0050),明胶酶谱检测的释放到胞外MMP-9活性明显高于对照组(P=0.0087),IL-6刺激后PMN中STAT3磷酸化水平明显升高(P=0.0089)。在IL-6作用时加入特异性STAT3抑制剂Ⅷ后,MMP-9 mRNA水平、培养上清液中MMP-9活性以及PMN中磷酸化STAT3蛋白水平与IL-6刺激组相比均明显下降(P=0.0067, 0.048和0.0098)。结论 IL-6刺激骨髓PMN中MMP-9的生成和释放,该作用可能是通过诱导STAT3信号通路磷酸化实现的。

     

    Abstract: Objective To investigate the effect of IL-6 inducing murine bone marrow neutrophils MMP-9 production and releasing and its mechanism of activation. Methods Murine bone marrow neutrophils were separated and cultured in vitro and randomly divided into four groups: control group, IL-6 50ng/ml stimulation group, IL-6 50ng/ml+DMSO group and IL-6 50ng/ml+STAT3 inhibitor Ⅷ 50μmol/L group. RT-PCR method was used to evaluate the mRNA level of intracellular MMP-9 of neutrophils. Gelatin zymography was performed to detect the activity of MMP-9 released from neutrophils. Western blot was applied to test the phosphorylation levels of STAT3 pathway in neutrophils. Results Compared with control group, the administration of IL-6 enhanced the mRNA expression of MMP-9(P=0.0050); the activity of MMP-9 released from neutrophils was elevated(P=0.0087) and the p-STAT3 levels in neutrophils was also increased (P=0.089). After the STAT3 inhibitor Ⅷ was added, the mRNA expression of MMP-9, the activities of MMP-9 in the supernatant and the p-STAT3 levels in neutrophils were decreased significantly than those in the IL-6 group (P=0.0067, 0.048 and 0.098, respectively). Conclusion IL-6 induces the production and releasing of MMP-9 in neutrophils and the effect may be induced by the activation of STAT3 pathway.

     

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出版历程
  • 收稿日期:  2015-04-14
  • 修回日期:  2015-11-19
  • 刊出日期:  2016-02-24

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