Effect of High MiR-146a Expression on the Inflammatory Reaction in BV2 Cells

Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2016 Feb;38(1):27-32. doi: 10.3881/j.issn.1000-503X.2016.01.005.

Abstract

Objective: To explore the effect of MiR-146a regulator function on the inflammatory response in neuroglia cell (microglia).

Methods: BV2 cells were transfected by MiR-146a mimics,and then stimulated by lipopolysaccharide (LPS). MiR-146a expression was measured by real-time polymerase chain reaction (real-time PCR). Interleukin (IL)-6 and tumor necrosis factor α (TNFα) were measured by enzyme-linked immunosorbent assay (ELISA). Furthermore, IL-1 receptor-associated kinase 1 (IRAK1) and TNF receptor-associated factor 6 (TRAF6) were detected by PCR and Western blotting.

Results: Compared to the normal control group, MiR-146a expression was significantly elevated by transfection with MiR-146a mimics (t=5.846, P=0.0021). The expression levels of IRAK1, TRAF6, TNFα, and IL-6 significantly increased in the LPS-stimulated BV2 cells compared to the non-stimulated BV2. The enhancement of MiR-146a resulted in significantly decreased IL-6 (t=5.200, P=0.0003) and TNFα (t=9.812, P<0.0001) secretion. The mRNA (t=5.353, P=0.0007) and protein (t=6.980, P=0.0009) levels of TRAF6, but not IRAK1, also significantly decreased.

Conclusion: MiR-146a may negatively suppress the inflammatory response of BV2 cells by regulating the expression of IRAF6 molecules in the TLR4 signaling pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • Cell Line
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Inflammation*
  • Interleukin-1 Receptor-Associated Kinases
  • Interleukin-6
  • Lipopolysaccharides
  • MicroRNAs
  • RNA, Messenger
  • Real-Time Polymerase Chain Reaction
  • Signal Transduction
  • TNF Receptor-Associated Factor 6
  • Transfection
  • Tumor Necrosis Factor-alpha

Substances

  • IL6 protein, human
  • Interleukin-6
  • Lipopolysaccharides
  • MicroRNAs
  • RNA, Messenger
  • TNF Receptor-Associated Factor 6
  • Tumor Necrosis Factor-alpha
  • IRAK1 protein, human
  • Interleukin-1 Receptor-Associated Kinases