Department of Respiration, Liaocheng People's Hospital, PR China.
Innate Immun. 2019 Oct;25(7):433-443. doi: 10.1177/1753425919861427. Epub 2019 Jul 10.
To investigate the role of miR-146 and its possible relationship with MALAT1 in LPS-induced inflammation in human microvascular endothelial cells (HMECs), HMEC-1 cells were treated with LPS to construct an inflammatory injury cell model, and the cell viability, TNF-α and IL-6 secretion and the expression levels of VCAM-1, SELE and ICAM-1 were analysed as markers of inflammatory injury. The regulation mechanisms of miR-146 interacted with MALAT1 and the downstream NF-κB signalling were also verified by dual-luciferase assay and knockdown technology. LPS significantly decreased the cell viability, increased levels of VCAM-1, SELE and ICAM-1 and also up-regulated miR-146a/b, TNF-α and IL-6 in a dose-dependent manner. Over-expression of miR-146a resulted in down-regulation of TNF-α and IL-6, as well as VCAM-1, SELE and ICAM-1, while inhibition of miR-146a led to opposite results. The dual-luciferase reporter assay showed both miR-146a and miR-146b directly targeted and negatively regulated the expression of MALAT1. Silencing of MALAT1 suppressed LPS-induced NF-κB activation and TNF-α and IL-6 secretion, reducing the cell inflammatory injury, but these changes were reversed after combined treatment with miR-146a inhibitor. Taken together, we demonstrate that miR-146 protects HMECs against inflammatory injury by inhibiting NF-κB activation. This process is modulated by MALAT1.
为了研究 miR-146 的作用及其与 MALAT1 之间可能的关系在 LPS 诱导的人微血管内皮细胞（HMEC）炎症中的作用，用 LPS 处理 HMEC-1 细胞构建炎症损伤细胞模型，分析细胞活力、TNF-α 和 IL-6 的分泌以及 VCAM-1、SELE 和 ICAM-1 的表达水平作为炎症损伤的标志物。通过双荧光素酶报告基因实验和敲低技术验证了 miR-146 与 MALAT1 相互作用的调控机制及其下游 NF-κB 信号通路。LPS 显著降低细胞活力，呈剂量依赖性增加 VCAM-1、SELE 和 ICAM-1 的水平，并上调 miR-146a/b、TNF-α 和 IL-6。miR-146a 的过表达导致 TNF-α 和 IL-6 以及 VCAM-1、SELE 和 ICAM-1 的下调，而 miR-146a 的抑制则导致相反的结果。双荧光素酶报告基因实验表明，miR-146a 和 miR-146b 均可直接靶向并负调控 MALAT1 的表达。沉默 MALAT1 抑制 LPS 诱导的 NF-κB 激活和 TNF-α 和 IL-6 的分泌，减轻细胞炎症损伤，但在与 miR-146a 抑制剂联合处理后，这些变化发生逆转。综上所述，我们证明 miR-146 通过抑制 NF-κB 激活来保护 HMEC 免受炎症损伤。这一过程受 MALAT1 调节。
