MicroRNA-146a inhibits NF-κB activation and pro-inflammatory cytokine production by regulating IRAK1 expression in THP-1 cells
MicroRNA-146a inhibits NF-κB activation and pro-inflammatory cytokine production by regulating IRAK1 expression in THP-1 cells
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MicroRNA-146a 通过调节 THP-1 细胞中 IRAK1 的表达来抑制 NF-kappa B 激活和促炎细胞因子的产生
DOI：
10.3892/etm.2019.7881
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发表时间：
2019-10-01
影响因子：
2.7
通讯作者：
Mu, Hong
中科院分区：
文献类型：
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作者：
Zhou, Chunlei;Zhao, Lan;Mu, Hong
系统性红斑狼疮（SLE）患者外周血单个核细胞中microRNA（miR）-146a水平降低;然而，其功能尚不清楚。本研究探讨了miR-146 a在脂多糖（LPS）诱导的THP-1细胞炎症中的调节作用。miR-146 a模拟物和抑制剂分别用于过表达和下调miR-146 a表达。逆转录定量PCR和蛋白质印迹分析，以评估白细胞介素（IL）-1受体相关激酶1（IRAK 1）的表达，蛋白质印迹分析，以评估核因子-κ B B激活，通过分析在核中的p65亚基水平。为了研究miR-146 a对LPS诱导的炎症的影响，还使用ELISA测量了IL-6和肿瘤坏死因子-α（TNF-alpha）水平。本研究的结果显示，miR-146 a过表达显著降低了LPS处理后THP-1细胞的IRAK 1表达，降低了细胞核中的p65水平，并降低了细胞培养基上清液中的IL-6和TNF-α水平。荧光素酶测定证实IRAK 1是THP-1细胞中miR-146 a的直接靶点。结论：miR-146 a可能通过调控IRAK 1的表达，抑制炎症信号的激活和促炎细胞因子的分泌。本研究至少部分揭示了miR-146 a调节SLE炎症反应的机制。
MicroRNA (miR)-146a levels are reduced in peripheral blood mononuclear cells of patients with systemic lupus erythematosus (SLE); however, its function is not well understood. The present study investigated the role of miR-146a in the regulation of lipopolysaccharide (LPS)-induced inflammation in THP-1 cells. A miR-146a mimic and an inhibitor were used to overexpress and downregulate miR-146a expression, respectively. Reverse transcription-quantitative PCR and western blot analyses were performed to evaluate interleukin (IL)-1 receptor-associated kinase 1 (IRAK1) expression, and western blot analysis was applied to assess nuclear factor-kappa B activation by analyzing p65 subunit levels in the nucleus. To investigate the effects of miR-146a on LPS-induced inflammation, IL-6 and tumor necrosis factor-alpha (TNF-alpha) levels were also measured using ELISA. The results of the present study revealed thatmiR-146a overexpression significantly reduced IRAK1 expression, reduced p65 levels in the nucleus and reduced IL-6 and TNF-alpha levels in the supernatant of the cell culture medium of THP-1 cells following LPS treatment. Luciferase assays confirmed IRAK1 to be a direct target of miR-146a in THP-1 cells. In conclusion, miR-146a may regulate IRAK1 expression and inhibit the activation of inflammatory signals and secretion of pro-inflammatory cytokines. The present study revealed, at least in part, the mechanisms by which miR-146a regulate the inflammatory response in SLE.