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Published on: August 23, 2019
miR-140-3p suppresses the proliferation and migration of macrophages
Pingping Qiao1, Jun Zhu2, Xiaoheng Lu3
1Nantong University, Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Co-innovation Center of Neuroregeneration, NMPA Key Laboratory for Research and Evaluation of Tissue Engineering Technology Products, Nantong, Jiangsu, China.
Abstract:
Macrophages benefit myelin debris removal, blood vessel formation, and Schwann cell activation following peripheral nerve injury. Identifying factors that modulate macrophage phenotype may advantage the repair and regeneration of injured peripheral nerves. microRNAs (miRNAs) are important regulators of many physiological and pathological processes, including peripheral nerve regeneration. Herein, we investigated the regulatory roles of miR-140-3p, a miRNA that was differentially expressed in injured rat sciatic nerves, in macrophage RAW264.7 cells. Observations from EdU proliferation assay demonstrated that elevated miR-140-3p decreased the proliferation rates of RAW264.7 cells while suppressed miR-140-3p increased the proliferation rates of RAW264.7 cells. Transwell-based migration assay showed that up-regulated and down-regulated miR-140-3p led to elevated and reduced migration abilities, respectively. However, the abundances of numerous phenotypic markers of M1 and M2 macrophages were not significantly altered by miR-140-3p mimic or inhibitor transfection. Bioinformatic analysis and miR-140-3p-induced gene suppression examination suggested that Smad3 might be the target gene of miR-140-3p. These findings illuminate the inhibitory effects of miR-140-3p on the proliferation and migration of macrophages and contribute to the cognition of the essential roles of miRNAs during peripheral nerve regeneration.
Insights
MicroRNAs (miRNAs) regulate macrophage behavior after nerve injury. This study shows miR-140-3p inhibits macrophage proliferation and migration, impacting peripheral nerve regeneration.
Area of Science:
- Neuroscience
- Molecular Biology
- Cell Biology
Background:
- Macrophages are crucial for peripheral nerve repair, aiding in debris clearance, new blood vessel formation, and Schwann cell activation.
- MicroRNAs (miRNAs) are key regulators in biological processes, including peripheral nerve regeneration.
- Understanding factors that influence macrophage behavior is vital for enhancing nerve repair strategies.
Purpose of the Study:
- To investigate the regulatory role of miR-140-3p, a differentially expressed miRNA in injured rat sciatic nerves, on macrophage RAW264.7 cells.
- To elucidate the impact of miR-140-3p on macrophage proliferation, migration, and phenotype.
- To identify potential molecular targets of miR-140-3p in the context of macrophage function.
Main Methods:
- Cell proliferation was assessed using EdU assays.
- Cell migration was evaluated using Transwell assays.
- Macrophage phenotype markers (M1/M2) were analyzed after miRNA transfection.
- Bioinformatic analysis and gene suppression assays were employed to identify miRNA targets.
Main Results:
- Elevated miR-140-3p levels decreased RAW264.7 cell proliferation, while suppressed levels increased it.
- Upregulated miR-140-3p enhanced macrophage migration, whereas downregulated miR-140-3p reduced migration.
- miR-140-3p did not significantly alter the expression of M1 and M2 macrophage phenotypic markers.
- Smad3 was identified as a potential target gene of miR-140-3p.
Conclusions:
- miR-140-3p plays an inhibitory role in macrophage proliferation and migration.
- These findings highlight the significant role of miRNAs in regulating macrophage functions during peripheral nerve regeneration.
- Targeting miR-140-3p may offer a therapeutic avenue for modulating macrophage activity in nerve repair.
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