基础医学与临床 ›› 2021, Vol. 41 ›› Issue (3): 404-408.

• 研究论文 • 上一篇    下一篇

Lnc-MALAT1减轻miR-217抑制LPS诱导的大鼠肺泡巨噬细胞炎性反应

罗传皊1, 孙利平2, 戚成栋1*   

  1. 枣庄矿业集团中心医院 1.重症医学科; 2.小儿内科, 山东 枣庄 277100
  • 收稿日期:2020-04-17 修回日期:2020-07-31 出版日期:2021-03-05 发布日期:2021-03-01
  • 通讯作者: *408978399@qq.com

Lnc-MALAT1 attenuates miR-217 inhibiting LPS-induced inflammatory response in rat alveolar macrophages

LUO Chuan-ling1, SUN Li-ping2, QI Cheng-dong1*   

  1. 1. Department of Critical Medicine; 2. Department of Pediatric Medicine, Zaozhuang Mining Group Central Hospital, Zaozhuang 277100,China
  • Received:2020-04-17 Revised:2020-07-31 Online:2021-03-05 Published:2021-03-01
  • Contact: *408978399@qq.com

摘要: 目的 探究长链非编码RNA 转移相关肺腺癌转录本1 (lnc-MALAT1)在脂多糖(LPS)诱导的大鼠肺泡巨噬细胞系(AMOs)炎性反应中的调控机制。方法 LPS处理AMOs建立细胞损伤模型,用脂质体法分别将pcDNA、pcDNA-MALAT1、miR-NC、miR-217 mimics、pcDNA-MALAT1与miR-NC、pcDNA-MALAT1与miR-217 mimics转染至AMOs,LPS处理12 h。采用酶联免疫吸附法(ELISA)检测细胞分泌人白介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)的量;采用RT-qPCR检测细胞中MALAT1、miR-217的表达;采用双荧光素酶报告实验、RNA结合蛋白免疫沉淀实验(RIP)检测MALAT1与miR-217的靶向关系。结果 与AMOs组相比,LPS+AMOs组细胞中IL-1β、TNFα的含量显著升高(P<0.05),MALAT1表达水平显著升高,miR-217表达水平显著降低(P<0.05)。双荧光素酶报告实验与RIP实验证实MALAT1能够靶向结合miR-217。过表达MALAT1可明显促进LPS诱导的AMOs细胞IL-1β、TNF-α的分泌,而过表达miR-217则可抑制LPS诱导的AMOs细胞IL-1β、TNF-α的分泌,并且过表达miR-217减轻MALAT1促进LPS诱导的AMOs细胞IL-1β、TNF-α的分泌。结论 Lnc-MALAT1可促进LPS诱导的AMOs的炎性反应,其机制与靶向miR-217相关。

关键词: lnc-MALAT1, 肺泡巨噬细胞, miR-217, 脂多糖(LPS)

Abstract: Objective To investigate the regulatory mechanism of long-chain noncoding RNA metastasis-associated lung adenocarcinoma 1 (lnc-MALAT1) in the inflammatory response of lipopolysaccharide (LPS)-induced alveolar macrophages (AMOs). Methods AMOs cells were treated with LPS for 12 hours to create an inflammation model, and liposome method was used to transfect pcDNA, pcDNA-MALAT1, miR-NC, miR-217 mimics, pcDNA-MALAT1 and miR-NC, pcDNA-MALAT1 and miR-217 mimics to AMOs. The level of IL-1β and TNF-α was detected by ELISA. RT-qPCR was used to detect the expression of MALAT1 and miR-217 in cells. The dual luciferase report experiment and RIP experiment were used to detect potential target-attacking relationship between MALAT1 and miR-217. Results Compared with the AMOs group, the expression of IL-1β and TNFα, MALAT1 in the cells of the LPS+AMOs group were significantly increased (P<0.05) and the expression level of miR-217 was significantly reduced(P<0.05). The dual luciferase report experiment and RIP experiment confirmed that MALAT1 targeted at miR-217. Over-expression of MALAT1 could obviously promote the secretion of IL-1β and TNF-α induced by LPS in AMOs, while over-expression of miR-217 inhibited the secretion of IL-1β and TNF-α induced by LPS in AMOs. Furthermore, over-expression of miR-217 reduced the secretion of IL-1β and TNF-α induced by MALAT1 in LPS-induced AMOs. Conclusions Lnc-MALAT1, a long non-coding RNA, can promote the inflammatory response of AMOs induced by LPS, and its mechanism is related to its targeting at miR-217.

Key words: lnc-MALAT1, alveolar macrophages, miR-217, lipopolysaccharide(LPS)

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