Background: As a key substance for intercellular communication, exosomes could be a potential strategy for stroke treatment. Activated microglia disrupt the integrity of blood - brain barrier (BBB) to facilitate the stroke process. Hence, this study was designed to investigate the effect of microglia-derived exosomes on BBB cell model injury and to explore the underlying molecular mechanisms. Methods: M1 polarization of BV2 cells was induced with LPS and their derived exosomes were isolated. Astrocytes were cultured in primary culture and constructed with End3 cells as a BBB cell model. After co-culture with exosomes, the BBB cell model was examined for changes in TEER, permeability, and expression of BBB-related proteins (Claudin-1, Occludin, ZO-1 and JAM). Resting and M1 -type BV2 cell-derived exosomes perform small RNA sequences and differentially expressed miRNAs (DE-miRNAs) are identified by bioinformatics. Results: M1 -type BV2 cell-derived exosomes decreased End3 cell viability, and increased their apoptotic ratio. Moreover, M1 type BV2 cell-derived exosomes dramatically enhanced the permeability of BBB cell model, and diminished the TEER and BBB-related protein (Claudin-1, Occludin, ZO-1) expression. Notably, resting BV2 cellderived exosomes had no effect on the integrity of BBB cell model. Sequencing results indicated that 71 DEmiRNAs were present in M1 BV2 cell-derived exosomes, and their targets mediated neurological development and signaling pathways such as MAPK and cAMP. RT-qPCR confirmed the differential expression of mmu-miR125a-5p, mmu-miR-122b-3p, mmu-miR-139-3p, mmu-miR-330-3p, mmu-miR-3057-5p and mmu-miR-342-3p consistent with the small RNA sequence. Furthermore, Creb1, Jun, Mtor, Frk, Pabpc1 and Sdc1 are the most well-connected proteins in the PPI network. Conclusion: M1 -type microglia-derived exosomes contribute to the injury of BBB cell model, which has the involvement of miRNAs. Our findings provide new perspectives and potential mechanisms for future M1 microglia-derived exosomes as therapeutic targets in stroke.
[1]Kunming Med Univ, Dept Neurol, Affiliated Hosp 1, 295 Xichang Rd, Kunming 650032, Yunnan, Peoples R China.
[2]Yunnan Prov Clin Res Ctr Neurol Dis, 295 Xichang Rd, Kunming 650032, Yunnan, Peoples R China.
[3]Kunming Med Univ, Dept Pain Med, Affiliated Hosp 1, 295 Xichang Rd, Kunming 650032, Yunnan, Peoples R China.
[4]Kunming Med Univ, Dept Cardiac Surg, Affiliated Hosp 1, 295 Xichang Rd, Kunming 650032, Yunnan, Peoples R China.
(8.0+极速模式)