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Updated: Mar 29, 2026
Primary Microglia Isolation from Mixed Glial Cell Cultures of Neonatal Rat Brain Tissue
Published on: August 15, 2012
NDRG2 promoted secreted miR-375 in microvesicles shed from M1 microglia, which induced neuron damage
Li-li Tang1, Yuan-bo Wu2, Chuan-qin Fang1
1Department of Neurology, the Second Affiliated Hospital of Anhui Medical University, Hefei, 230601, China.
Background:
Microglia microvesicles (MVs) has shown to have significant biological functions under normal conditions. A diversity of miRNAs is involved in neuronal development, survival, function, and plasticity, but the exact functional role of NDRG2 and secreted miR-375 in MVs in neuron damage is poorly understood. We investigated the effect of NDRG2 and secreted miR-375 in MVs shed from M1 microglia on neuron damage.
Methods:
Expression of Nos2, Arg-1, miR-375, syntaxin-1A, NDRG2 and Pdk 1 were evaluated using RT-PCR or western blotting. Cell viability of N2A neuron was quantified by a MTT assay.
Results:
Microglia can be polarized into different functional phenotypes. Expression of NDRG2 and Nos2 were significantly increased by LPS treatment on N9 cells, whereas treatment with IL-4 dramatically suppressed the expression of NDRG2 and remarkably elevated expression of Arg-1. Besides, MVs shed from LPS-treated N9 microglia significantly inhibited cell viability of N2A neurons and expression of syntaxin-1A, and NDRG2 interference reversed the up-regulated miR-375 in LPS-treated N9 microglia and MVs shed from LPS-treated N9 cells. Furthermore, NDRG2 could modulate miR-375 expression in N9 microglia and MVs. And miR-375 inhibitor remarkably elevated Pdk1 expression in N2A neurons. Finally, miR-375 inhibitor could reverse suppression effect of NDRG2 overexpression on cell viability of N2A neurons and expression of syntaxin-1A.
Conclusion:
Our results demonstrated that NDRG2 promoted secreted miR-375 in microvesicles shed from M1 microglia, which induced neuron damage. The suppression of NDRG2 and secreted miR-375 in MVs shed from M1 microglia may be potential targets for alleviation of neuron damage.
Insights
NDRG2 promotes neuron damage by increasing miR-375 in microglia microvesicles. Suppressing NDRG2 and miR-375 in MVs may help alleviate neuron damage.
Area of Science:
- Neuroscience
- Cell Biology
- Molecular Biology
Background:
- Microglia microvesicles (MVs) have crucial roles in biological functions.
- The roles of NDRG2 and miR-375 in MVs concerning neuron damage are not fully understood.
Purpose of the Study:
- To investigate the effect of NDRG2 and secreted miR-375 in MVs from M1 microglia on neuron damage.
Main Methods:
- Utilized RT-PCR and western blotting to assess gene and protein expression (Nos2, Arg-1, miR-375, syntaxin-1A, NDRG2, Pdk 1).
- Quantified N2A neuron viability using MTT assay.
- Manipulated microglia polarization and gene expression to study molecular mechanisms.
Main Results:
- LPS treatment increased NDRG2 and Nos2 in N9 cells; IL-4 had opposite effects.
- MVs from LPS-treated microglia reduced N2A neuron viability and syntaxin-1A expression.
- NDRG2 interference reversed miR-375 upregulation in microglia and MVs; NDRG2 modulated miR-375 expression.
- miR-375 inhibition increased Pdk1 in neurons and reversed NDRG2's suppressive effects on neuron viability and syntaxin-1A.
Conclusions:
- NDRG2 promotes neuron damage by enhancing miR-375 secretion in MVs from M1 microglia.
- Targeting NDRG2 and miR-375 in MVs presents a potential therapeutic strategy for neuron damage.
