M1 macrophage-derived exosomal microRNA-22-3p regulates vascular endothelial senescence
OUYANG Shun
LI Wenling
QIU Yumin
LIU Zhangchi
ZHU Qiuxia
ZHANG Meixin
XU Lingling
TAO Jun
Abstract：Objective To investigate the role of M1 macrophage-derived exosomes(M1-Exo)in regulating vascular endothelial senescence and explore the related molecular mechanism.Methods Six young mice and 6 aged mice were randomly subjected and assigned into a young group and an aged group.The flow-mediated dilation(FMD)of the brachial artery was compared between the two groups.Endothelial cells were co-cultured with solvent control(solvent group),100 ng/μL M1-Exo(M1-Exo 100 ng/μL group),and 200 ng/μL M1-Exo(M1-Exo 200 ng/μL group)respectively.Senescence-associatedβ-galactosidase(SA-β-gal)staining was carried out,and confocal imaging was used to detect the production of intracellular mitochondrial reactive oxygen species(ROS).The following groups were established:microRNA(miRNA)blank control group,miR-22-3p mimic group,miRNA negative control group,and miR-22-3p inhibitor group.The protein expression of silent mating type information regulation 2 homolog-1(SIRT1)was compared among these above groups.Short hairpin RNA(shRNA)transfection was performed with shRNA-SIRT1 plasmid to establish a sh-SIRT1 group,and the cells transfected with shRNA blank plasmid served as control group(sh-NC group).Western blotting was used to detect protein expression in both groups.Results The FMD was significantly decreased in the aged group than the young group[(48.330±3.807)％ vs(28.650±3.209)％,P＜0.05].In vitro experiments showed that compared with solvent group,the SA-β-gal expression and mitochondrial ROS levels were elevated in both M1-Exo 100 ng/μL and M1-Exo 200 ng/μL groups(P＜0.05).The SIRT1 level was decreased in the miR-22-3p mimic group than the miRNA blank control group,but increased in the miR-22-3p inhibitor group than the miR-22-3p mimic group(P＜0.05).Compared with the sh-NC group,the nuclear factor κB/cyclin-dependent kinase inhibitor P21(NF-κB/P21)and Forkhead box O3α/superoxide dismutase 2(FOXO3α/SOD2)signaling pathways were activated in the sh-SIRT1 group.Conclusion M1 macrophage-derived exosomal miR-22-3p promotes vascular endothelial cell senescence.
Keywords：endothelial cellsagingexosomesinflammationmacrophagesmicroRNAs
Publication Date：2026-03-15
Online Publishing Date：2026-03-31(First online date of this platform, not the publication date of the document)
Pages：7( 294-300 )