Department of Plastic and Burn, Baoji Center Hospital, Baoji, Shaanxi 721008, P.R. China.
Clinical Laboratory, Baoji Center Hospital, Baoji, Shaanxi 721008, P.R. China.
Mol Med Rep. 2019 May;19(5):3756-3766. doi: 10.3892/mmr.2019.10028. Epub 2019 Mar 14.
The present study aimed to identify the involvement of the dysregulation of the metastasis‑associated lung adenocarcinoma transcript‑1 (MALAT1)/microRNA (miR)‑214/Toll‑like receptor (TLR)5 signaling pathway in the development of post‑burn sepsis. THP‑1 cells were used in the present study, in addition to 8‑10 week‑old mice. ELISA analysis was performed to examine the expression levels of inflammation‑associated factors. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) analysis and western blotting were performed to analyze the influence of burns or burns with infection on the production of MALAT1, miR‑214 and TLR5. Commonly‑used software and a luciferase assay was used to confirm the target gene of miR‑214. RT‑qPCR analysis and western blotting were performed to elucidate the effects of lipopolysaccharide (LPS), miR‑214 and MALAT1 on the expression of miR‑214, TLR5, tumor necrosis factor (TNF)‑α, interleukin (IL)‑6 and IL‑10. Burn injury increased TLR5, TNF‑α, IL‑6 and IL‑10 expression levels, which were abolished by treatment with MALAT1. miR‑214 directly targeted TLR5 by binding to the TLR5 3' untranslated region (ITR), and the luciferase activity of the wild‑type, and not the mutant, TLR5 3'UTR was reduced following transfection with miR‑214. In cells not treated with LPS, MALAT1 and anti‑miR‑214 significantly enhanced TLR5, TNF‑α, IL‑6 and IL‑10 expression, and repressed miR‑214 production; whereas, miR‑214 and MALAT1 short hairpin (sh)RNA decreased TLR5, TNF‑α, IL‑6 and IL‑10 expression levels, and increased miR‑214 expression. In cells treated with LPS, LPS reduced miR‑214 expression and increased TLR5, TNF‑α, IL‑6 and IL‑10 expression compared with LPS‑untreated cells, and the effects of MALAT1, anti‑miR‑214, miR‑214 and MALAT1 shRNA on TLR5, TNF‑α, IL‑6 and IL‑10 were the same as in LPS‑untreated cell. The results of the present study indicated the association between the dysregulation of MALAT1/miR‑214/TLR5 and the risk of post‑burn sepsis.
本研究旨在确定失调的转移相关肺腺癌转录物 1（MALAT1）/microRNA（miR）-214/Toll 样受体（TLR）5 信号通路在烧伤后脓毒症发展中的作用。本研究使用了 THP-1 细胞和 8-10 周龄的小鼠。采用 ELISA 分析检测炎症相关因子的表达水平。采用逆转录-定量聚合酶链反应（RT-qPCR）分析和 Western blot 分析检测烧伤或烧伤合并感染对 MALAT1、miR-214 和 TLR5 产生的影响。采用常用软件和荧光素酶检测实验证实 miR-214 的靶基因。采用 RT-qPCR 分析和 Western blot 分析阐明脂多糖（LPS）、miR-214 和 MALAT1 对 miR-214、TLR5、肿瘤坏死因子（TNF）-α、白细胞介素（IL）-6 和 IL-10 表达的影响。烧伤增加了 TLR5、TNF-α、IL-6 和 IL-10 的表达水平，而 MALAT1 的处理则消除了这种表达。miR-214 通过与 TLR5 3'非翻译区（UTR）结合直接靶向 TLR5，并且转染 miR-214 后野生型而非突变型 TLR5 3'UTR 的荧光素酶活性降低。在未用 LPS 处理的细胞中，MALAT1 和抗 miR-214 显著增强了 TLR5、TNF-α、IL-6 和 IL-10 的表达，并抑制了 miR-214 的产生；而 miR-214 和 MALAT1 短发夹（sh）RNA 则降低了 TLR5、TNF-α、IL-6 和 IL-10 的表达水平，并增加了 miR-214 的表达。在 LPS 处理的细胞中，与未用 LPS 处理的细胞相比，LPS 降低了 miR-214 的表达并增加了 TLR5、TNF-α、IL-6 和 IL-10 的表达，MALAT1、抗 miR-214、miR-214 和 MALAT1 shRNA 对 TLR5、TNF-α、IL-6 和 IL-10 的影响与未用 LPS 处理的细胞相同。本研究结果表明 MALAT1/miR-214/TLR5 失调与烧伤后脓毒症风险之间存在关联。
