基础医学与临床 ›› 2017, Vol. 37 ›› Issue (9): 1237-1242.

• 研究论文 • 上一篇    下一篇

α7烟碱型乙酰胆碱受体激动剂抑制骨水泥微粒刺激小鼠外周血单核细胞分泌炎性因子

沈凯1,谭祖键1,杨阜滨1,张胜利1,曹代桂1,邓忠良2   

  1. 1. 重庆市人民医院骨科
    2. 重庆医科大学附属第二医院骨科
  • 收稿日期:2017-02-20 修回日期:2017-05-30 出版日期:2017-09-05 发布日期:2017-08-28
  • 通讯作者: 邓忠良 E-mail:yizhujuncqmu@163.com
  • 基金资助:
    IGF1对地塞米松抑制干细胞成骨分化的影响及机制研究

α7 Nicotinic acetylcholine receptor agonist inhibits bone cement particles stimulated secretion of inflammatory cytokines in peripheral blood monocytes of mice

  • Received:2017-02-20 Revised:2017-05-30 Online:2017-09-05 Published:2017-08-28

摘要: 目的 探讨α7烟碱型乙酰胆碱受体(α7nAChR)激动剂PNU282987对骨水泥微粒刺激小鼠外周血单核细胞分泌炎性反应因子的影响及其分子机制。方法 分离培养小鼠外周血单核细胞,使用聚甲基丙烯酸甲酯(PMMA)微粒刺激后,ELISA检测培养上清液中TNF-α、IL-1β和IL-6的含量;RT-PCR检测细胞TNF-α、IL-1β和IL-6的mRNA表达;Western blot检测p-p65、p65、p-JAK2、JAK2、p-stat3、stat3及β-actin的表达;ELISA检测NF-?B DNA结合活力。 结果 单核细胞经PMMA微粒刺激后,上清液中TNF-α、IL-1β和IL-6的含量明显增高(P<0.05);细胞TNF-α、IL-1β和IL-6 mRNA表达明显增高(P<0.05);p65、JAK2和stat 3磷酸化明显增强(P<0.05);NF-κB DNA结合活力明显增高(P<0.05)。不同浓度PNU282987作用后,上清液中TNF-α、IL-1β和IL-6的含量呈浓度依耐性下降(P<0.05);细胞TNF-α、IL-1β和IL-6 mRNA表达呈浓度依耐性下降(P<0.05);总p65、JAK2和stat3的表达不变;p-p65、p-JAK2和p-stat3的表达呈浓度依耐性下降(P<0.05);NF-κB DNA结合活力也呈浓度依耐性下降(P<0.05)。 结论 α7nAChR激动剂PNU282987能显著抑制PMMA骨水泥微粒所诱导的小鼠血单核细胞炎性反应因子的分泌。

关键词: α7烟碱型乙酰胆碱受体, 单核细胞, 核因子κB, 信号传导与转录激活因子3

Abstract: Objective To investigate the effect of α7 nicotinic acetylcholine receptor (α7nAChR) agonists agent PNU282987 on bone cement particles stimulated secretion of inflammatory cytokines in blood monocytes and its molecular mechanism. Methods Mouse peripheral blood monocytes were isolated and the inflammatory response were induced by PMMA particles. TNF-α, IL-1β and IL-6 concentration in culture supernatant were measured by ELISA. TNF-α, IL-1β and IL-6 mRNA expression were measured by RT-PCR. P-p65, p65, p-JAK2, JAK2, p-STAT3, STAT3, and β-actin expression were detected by Western blot. NF-?B DNA binding activity were measured by ELISA. Results After stimulation of PMMA particles, TNF-α, IL-1β and IL-6 concentration in culture supernatant were significantly increased (P<0.05), TNF-α, IL-1β and IL-6 mRNA expression were significantly increased (P<0.05), p-p65, p-JAK2 and p-STAT3 expression and NF-κB DNA binding activity were also increased significantly (P<0.05). However, after PNU282987 treatment, TNF-α, IL-1β and IL-6 concentration in culture supernatant were decreased in a concentration gradient way (P<0.05), TNF-α, IL-1β and IL-6 mRNA expression were decreased in a concentration gradient way (P<0.05), p-p65, p-JAK2 and p-STAT3 expression and NF-κB p65 DNA binding activity were also decreased in a concentration gradient way (P<0.05). Conclusion A7nAChR agonist PNU282987 significantly inhibited PMMA bone cement particles induced secretion of inflammatory cytokines in blood mononuclear cells of mice.

Key words: α7nAChR, monocyte, NF-κB, STAT3

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