微小RNA-223的下调通过靶向肥大细胞中的胰岛素样生长因子1受体（IGF-1R），经由磷脂酰肌醇-3-激酶/蛋白激酶B（PI3K/Akt）信号通路促进脱颗粒作用。
Down-regulation of microRNA-223 promotes degranulation via the PI3K/Akt pathway by targeting IGF-1R in mast cells.
作者信息
Wang Quan, Zhao De-Yu, Xu Hong, Zhou Hui, Yang Qian-Yuan, Liu Feng, Zhou Guo-Ping
机构信息
Department of Respiratory Medicine, Nanjing Children's Hospital Affiliated to Nanjing Medical University, Nanjing, China.
Department of Pediatrics, Nanjing Maternal and Child Health Hospital of Nanjing Medical University, Nanjing, China.
出版信息
PLoS One. 2015 Apr 13;10(4):e0123575. doi: 10.1371/journal.pone.0123575. eCollection 2015.
BACKGROUND
Mast cells play a central role in allergic and inflammatory disorders by inducing degranulation and inflammatory mediator release. Recent reports have shown that miRNAs play an important role in inflammatory response regulation. Therefore, the role of miR-223 in mast cells was investigated.
METHODS
The expression of miR-223 was quantified by quantitative real-time polymerase chain reaction (qRT-PCR) in immunoglobulin E (IgE)-mediated mast cells. After successful miR-223 inhibition by transfection, degranulation was detected in IgE-mediated mast cells. The phosphorylation of IκB-α and Akt were examined using western blotting. NF-κB was tested using electrophoretic mobility shift assay. PI3K-inhibitor (LY294002) was used to investigate whether the PI3K/Akt pathway was essential for mast cell activation. The TargetScan database and a luciferase reporter system were used to identify whether insulin-like growth factor 1 receptor (IGF-1R) is a direct target of miR-223.
RESULTS
MiR-223 expression was up-regulated in IgE-mediated mast cells, whereas its down-regulation promoted mast cell degranulation. Levels of IκB-α and Akt phosphorylation as well as NF-κB were increased in miR-223 inhibitor cells. LY294002 could block the PI3K/Akt signaling pathway and rescue the promotion caused by suppressing miR-223 in mast cells. IGF-1R was identified as a direct target of miR-223.
CONCLUSIONS
These findings suggest that down-regulation of miR-223 promotes degranulation via the PI3K/Akt pathway by targeting IGF-1R in mast cells.
背景
肥大细胞通过诱导脱颗粒和炎症介质释放，在过敏性和炎症性疾病中发挥核心作用。最近的报道表明，微小RNA（miRNA）在炎症反应调节中起重要作用。因此，研究了miR-223在肥大细胞中的作用。
方法
通过定量实时聚合酶链反应（qRT-PCR）对免疫球蛋白E（IgE）介导的肥大细胞中miR-223的表达进行定量。转染成功抑制miR-223后，检测IgE介导的肥大细胞中的脱颗粒情况。使用蛋白质免疫印迹法检测IκB-α和Akt的磷酸化。使用电泳迁移率变动分析检测核因子κB（NF-κB）。使用磷脂酰肌醇-3激酶（PI3K）抑制剂（LY294002）研究PI3K/Akt信号通路对肥大细胞活化是否必不可少。使用TargetScan数据库和荧光素酶报告系统鉴定胰岛素样生长因子1受体（IGF-1R）是否是miR-223的直接靶标。
结果
在IgE介导的肥大细胞中，miR-223表达上调，而其下调促进肥大细胞脱颗粒。在miR-223抑制剂处理的细胞中，IκB-α和Akt的磷酸化水平以及NF-κB水平升高。LY294002可阻断PI3K/Akt信号通路，并挽救肥大细胞中抑制miR-223所导致的促进作用。IGF-1R被鉴定为miR-223的直接靶标。
结论
这些发现表明，在肥大细胞中，miR-223的下调通过靶向IGF-1R，经由PI3K/Akt信号通路促进脱颗粒。
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