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,孙利平,戚成栋   

  1. 枣庄矿业集团中心医院
  • 收稿日期:2020-04-17 修回日期:2020-07-28 出版日期:2021-03-05 发布日期:2021-03-01
  • 通讯作者: 罗传皊 E-mail:zx5fsa@163.com

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

  • Received:2020-04-17 Revised:2020-07-28 Online:2021-03-05 Published:2021-03-01

摘要: 目的 探究长链非编码RNA 转移相关肺腺癌转录本1 (LncMALAT1)在脂多糖(LPS)诱导的大鼠肺泡巨噬细胞(AMOs)炎性反应中的调控机制。 方法 LPS处理AMOs细胞建立细胞损伤模型,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(ELISA)法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的含量均显著升高(P<0.05),MALAT1表达显著升高,miR-217表达显著降低(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 长链非编码RNA MALAT1可促进LPS诱导的AMOs细胞的炎性反应,其机制与靶向miR-217相关。

关键词: LncMALAT1, 肺泡巨噬细胞, miR-217, 炎症

Abstract: Objective To investigate the regulatory mechanism of long-chain noncoding RNA metastasis-associated lung adenocarcinoma 1 (LncMALAT1) in the inflammatory response of lipopolysaccharide (LPS) -induced alveolar macrophages (AMOs). Methods LPS treated AMOs cells to establish a cell damage 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, LPS treatment for 12 h. The levels of IL-1β and TNF-α were detected by ELISA. RT-PCR 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 the targeting relationship between MALAT1 and miR-217. Results Compared with the AMOs group, the levels of IL-1β and TNFα in the cells of the LPS+AMOs group were significantly increased (P<0.05), the expression of MALAT1 was significantly increased, and the expression of miR-217 was significantly reduced (P<0.05). The dual luciferase report experiment and RIP experiment confirmed that MALAT1 could target and bind to miR-217. Overexpression of MALAT1 could obviously promote the secretion of IL-1β and TNF-α induced by LPS in AMOs, while overexpression of miR-217 could inhibit the secretion of IL-1β and TNF-α induced by LPS in AMOs. Furthermore, overexpression of miR-217 could reduce the secretion of IL-1β and TNF-α induced by MALAT1 in LPS-induced AMOs. Conclusions MALAT1, a long non-coding RNA, can promote the inflammatory response of AMOs cells induced by LPS, and its mechanism is related to targeting miR-217.

Key words: LncMALAT1, alveolar macrophages, miR-217, inflammation