Dataset Information

M2 microglia-derived exosomes inhibit p38 MAPK signaling pathways and GPX4-mediated neuronal ferroptosis in neuropathic pain [EXO_miRNAseq]

ABSTRACT: This study aimed to investigate the regulatory mechanism of M2 microglial exosomes (M2-exosomes) on neuronal ferroptosis during the progression of neuropathic pain (NP). IL-4 was used to induce M2 polarization of resting BV2 microglia. Subsequently, M2-exosomes were extracted from M2-polarized microglia and then sequenced. We further performed high-throughput sequencing for BV2 microglia and N2a neuronal cells that were cocultured with M2-exosomes. The effects of M2-exosomes on microglia and neuronal cells were detected by qRT-PCR, western blot, immunofluorescence, and flow cytometry in vitro. For the in vivo experiments, NP model mice were constructed using the chronic constriction sciatic nerve injury (CCI) method and injected intrathecally with M2-exosomes. Pain behavior and neuronal ferroptosis in the NP model mice were then assessed. The in vitro experiments indicated that M2-exosomes inhibited BV2 microglial inflammation response. The sequencing results for N2a neuronal cells treated with M2-exosomes showed that ferroptosis and the MAPK signaling pathway were significantly enriched. We further demonstrated that M2-exosomes reversed erastin-induced neuronal ferroptosis and inhibited activation of the p38 MAPK pathway in neuronal cells. Specifically, M2-exosomes increased expression of the ferroptosis antagonist factor GPX4 and decreased lipid peroxidation and mitochondrial ferrous levels in neuronal cells. The in vivo experiments using the CCI model showed that M2-exosomes increased GPX4 expression in spinal cord neuronal cells, inhibited the increase in ferrous content, and alleviated hyperalgesia. Our findings suggest that M2-exosomes activate the GPX4 antioxidant factor and inhibit p38 MAPK signaling pathways in neuronal ferroptosis, thereby improving pain hypersensitivity in NP.

ORGANISM(S): Mus musculus

PROVIDER: GSE306183 | GEO | 2026/08/21

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:Numerous neurological diseases involve neuroinflammation, a process in which immune cells, particularly microglia, contribute to neuronal death. Ferroptosis, a recently identified form of regulated cell death, is implicated in various diseases characterized by neuronal injury. Nicotinamide mononucleotide (NMN), a potent NAD+ precursor supplement, has been found to inhibit neuroinflammation and ferroptosis. However, the mechanisms of NMN in both ferroptosis and neuroinflammation remains unclear. The present study aimed to investigate the impact of NMN on neuroinflammation and the susceptibility of microglia to ferroptosis. Ferroptosis markers in microglia exposed to lipopolysaccharide (LPS) were analyzed using CCK8, flow cytometry, ELISA, and RT-qPCR. The effects of NMN on LPS-induced ferroptosis in microglia were evaluated through flow cytometry, western blotting, and immunofluorescence staining. RT-qPCR analysis assessed the inflammatory cytokine production of microglia subjected to ferrostatin-1-regulated ferroptosis. RNA sequencing elucidated the underlying mechanisms of NMN-associated microglia ferroptosis under LPS induction. In BV2 microglia, an inhibitor of Glutathione Peroxidase 4(GPX4), RSL3, was employed to suppress GPX4 expression. Intracerebroventricular injection of LPS was performed to evaluate neuroinflammation and microglia activation in vivo. LPS treatment resulted in decreased cell viability, accompanied by upregulation of ferroptosis markers SLC7A11 and GPX4, and elevated levels of malondialdehyde (MDA), 4-hydroxynonenal (4-HNE), and total iron in a dose-dependent manner. NMN effectively rescued LPS-induced ferroptosis and improved cell viability in microglia. Co-administration of NMN and ferrostatin-1 significantly reduced proinflammatory cytokine production in microglia following the introduction of LPS stimuli. Mechanistically, NMN facilitated glutathione (GSH) production, and promoted resistance to lipid peroxidation in a GPX4-dependent manner, repressing cytokine transcription and protecting cells from ferroptosis. RNA sequencing elucidated the underlying mechanism of NMN-associated microglia ferroptosis under LPS induction. Furthermore, simultaneous injection of NMN ameliorated LPS-induced ferroptosis and neuroinflammation in mouse brains. The data from the present study indicated that NMN enhances GPX4-mediated ferroptosis defense against LPS-induced ferroptosis in microglia by recruiting GSH, thereby inhibiting neuroinflammation. Therefore, therapeutic approaches targeting ferroptosis in diseases using NMN should consider both its anti-ferroptosis and anti-inflammatory effects to achieve optimal outcomes, presenting promising strategies for treating neuroinflammation-related diseases or disorders.

Dataset Information

M2 microglia-derived exosomes inhibit p38 MAPK signaling pathways and GPX4-mediated neuronal ferroptosis in neuropathic pain [EXO_miRNAseq]

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets