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Adipose-Derived Mesenchymal Stromal Cells Co-Cultured with Primary Mixed Glia to Reduce Prion-Induced Inflammation
Published on: August 11, 2023
MSC-derived exosomes delivery of miR-714 protect mouse BV2 microglial cells from OGD/R injury by directly targeting
Xiao-Xia Yang1, Yong-Jie Wang2, Ai-Ying Xue1
1Department of Pharmacy, The Second Qilu Hospital of Shandong University, Jinan, Shandong, 250033, PR China.
Abstract:
MicroRNAs have been widely demonstrated to participate in the pathogenesis of cerebrovascular diseases. In this study, an in vitro cellular oxygen-glucose deprivation/reoxygenation (OGD/R) model was established in mouse BV2 microglial cells, and differentially expressed microRNAs (DEMs) were screened by small RNA sequencing. The significant downregulation of miR-714 during OGD/R process was confirmed and its effects was verified by CCK-8 and apoptosis assays. MSC-derived exosomes (MSC-exo) were used to transport miR-714, and the mRNA expression level and cellular uptake of miR-714 were detected by qRT-PCR and fluorescence microscopy, respectively. The effects of MSC-exo-miR-714 on the proliferation and apoptosis of OGD/R BV2 cells were evaluated by CCK-8, Annexin V-FITC/PI double staining and TUNEL assays. In addition, bioinformatics analysis was performed to predict the target genes of miR-714. qRT-PCR and Western blotting were used to verify the regulatory effects of MSC-exo-miR-714 on ITGAM (integrin subunit alpha M) at mRNA and protein levels. The potential anti-inflammatory mechanism was further explored using gene enrichment analysis and Western blotting. The results showed that MSC-exo-miR-714 could significantly promote proliferation, inhibit apoptosis, and reduce protein expression of IL-6, IL-1β, TNF-α, NF-κB, and p-NF-κB in OGD/R-injured BV2 cells by directly targeting ITGAM. In conclusion, miR-714 is significantly down-regulated under in vitro OGD/R injury and serves as a key regulator in BV2 microglial injury. MSC-exo-miR-714 protects glial cells against OGD/R damage in vitro by directly targeting ITGAM and inhibiting inflammatory signaling. These findings are limited to the in vitro setting, and further in vivo validation is required.
Insights
MicroRNA-714, downregulated in oxygen-glucose deprivation/reoxygenation injury, protects microglial cells. Mesenchymal stem cell-derived exosomes carrying miR-714 promote cell proliferation and reduce inflammation by targeting ITGAM.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- MicroRNAs (miRNAs) play critical roles in the pathogenesis of cerebrovascular diseases.
- Microglial cells are central to neuroinflammation and injury following ischemic events.
Purpose of the Study:
- To investigate the role of miR-714 in oxygen-glucose deprivation/reoxygenation (OGD/R)-induced microglial injury.
- To evaluate the therapeutic potential of mesenchymal stem cell-derived exosomes (MSC-exo) delivering miR-714.
Main Methods:
- Established an in vitro OGD/R model using mouse BV2 microglial cells.
- Performed small RNA sequencing to identify differentially expressed miRNAs.
- Utilized CCK-8, apoptosis assays, qRT-PCR, and Western blotting to assess miR-714 function and its target ITGAM.
- Investigated the anti-inflammatory effects of MSC-exo-miR-714.
Main Results:
- miR-714 was significantly downregulated in OGD/R-injured BV2 cells.
- MSC-exo-miR-714 treatment promoted BV2 cell proliferation and inhibited apoptosis.
- MSC-exo-miR-714 directly targeted ITGAM, reducing pro-inflammatory mediators (IL-6, IL-1β, TNF-α) and NF-κB signaling.
Conclusions:
- miR-714 acts as a key regulator in microglial response to OGD/R injury.
- MSC-exo-miR-714 demonstrates protective effects against OGD/R-induced glial cell damage in vitro.
- Targeting ITGAM via exosomal miR-714 represents a potential therapeutic strategy for cerebrovascular injury, requiring further in vivo validation.
