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Published on: January 7, 2019
MiR-200c-3p inhibits LPS-induced M1 polarization of BV2 cells by targeting RIP2
Lei Zhao1, Xiaosong Liu1, Jiankai Yang1
1Department of Neurosurgery, The Second Hospital of Hebei Medical University, 215 Heping West Road, 050000, Shijiazhuang, Hebei, China.
Background:
Microglia are important immune cells, which can be induced by lipopolysaccharide (LPS) into M1 phenotype that express pro-inflammatory cytokines. Some studies have shown that microRNAs play critical roles in microglial activation.
Objective:
This study was designed to investigate the role of miR-200c-3p in regulating inflammatory responses of LPS-treated BV2 cells.
Methods:
The expression of miR-200c-3p in BV2 cells was detected by real-time PCR. Receptor-interacting protein 2 (RIP2) was predicted as a target gene of miR-200c-3p. Their relationship was verified by dual-luciferase reporter assay. The function of miR-200c-3p and RIP2 in microglial polarization and NF-κB signaling was further evaluated.
Results:
LPS treatment reduced miR-200c-3p expression in a dose-dependent and time-dependent manner in BV2 cells. LPS treatment increased the expression of M1 phenotype markers inducible nitric oxide synthase (iNOS) and major histocompatibility complex class (MHC)-II, promoted the release of pro-inflammatory cytokines interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α, and enhanced the nuclear translocation and phosphorylation of nuclear factor-kappaB (NF-κB) p65. Reversely, miR-200c-3p mimics down-regulated the levels of these inflammatory factors. Furthermore, RIP2 was identified to be a direct target of miR-200c-3p. RIP2 knockdown had a similar effect to miR-200c-3p mimics. Overexpression of RIP2 eliminated the inhibitory effect of miR-200c-3p on LPS-induced M1 polarization and NF-κB activation in BV2 cells.
Conclusions:
MiR-200c-3p mimics suppressed LPS-induced microglial M1 polarization and NF-κB activation by targeting RIP2. MiR-200c-3p/RIP2 might be a potential therapeutic target for the treatment of neuroinflammation-associated diseases.
Insights
MicroRNA miR-200c-3p suppresses M1 microglial polarization and inflammation by targeting RIP2. This miR-200c-3p/RIP2 pathway offers a potential therapeutic target for neuroinflammation.
Area of Science:
- Neuroimmunology
- Molecular Biology
Background:
- Microglia are key immune cells in the central nervous system.
- Lipopolysaccharide (LPS) induces microglia into a pro-inflammatory M1 phenotype.
- MicroRNAs are recognized regulators of microglial activation.
Purpose of the Study:
- To investigate the role of miR-200c-3p in LPS-induced inflammatory responses in BV2 microglial cells.
- To elucidate the regulatory mechanism of miR-200c-3p in microglial polarization.
Main Methods:
- Quantitative real-time PCR to detect miR-200c-3p expression.
- Dual-luciferase reporter assay to confirm RIP2 as a direct target of miR-200c-3p.
- Evaluation of microglial polarization markers, inflammatory cytokine release, and NF-κB signaling pathway activation.
Main Results:
- LPS treatment decreased miR-200c-3p expression in BV2 cells.
- LPS increased M1 markers (iNOS, MHC-II), pro-inflammatory cytokines (IL-1β, IL-6, TNF-α), and NF-κB activation.
- miR-200c-3p mimics reversed LPS-induced M1 polarization and inflammation.
- RIP2 was confirmed as a direct target; RIP2 knockdown mimicked miR-200c-3p effects, while RIP2 overexpression counteracted them.
Conclusions:
- miR-200c-3p inhibits LPS-induced microglial M1 polarization and NF-κB activation by targeting RIP2.
- The miR-200c-3p/RIP2 axis represents a potential therapeutic target for neuroinflammation-associated diseases.
